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Article

Diversity, Succession and Seasonal Variation of Phylloplane Mycoflora of Leucaena leucocephala in Relation to Its Leaf Litter Decomposition

1
Department of Botany, Dyal Singh College, University of Delhi, Delhi 110003, India
2
Department of Zoology, Kalindi College, University of Delhi, Delhi 110008, India
3
Department of Zoology, Ramjas College, University of Delhi, Delhi 110007, India
4
Department of Botany, SGTB Khalsa College, University of Delhi, Delhi 110007, India
*
Author to whom correspondence should be addressed.
J. Fungi 2022, 8(6), 608; https://doi.org/10.3390/jof8060608
Submission received: 15 May 2022 / Revised: 2 June 2022 / Accepted: 4 June 2022 / Published: 6 June 2022

Abstract

:
To address international food security concerns and sustain a growing global population, global agricultural output needs to increase by 70% by the year 2050. Current agricultural techniques to increase crop yields, specifically the application of chemicals, have resulted in a wide range of negative impacts on the environment and human health. The maintenance of good quality soil organic matter, a key concern in tropical countries such as India, requires a steady input of organic residues to maintain soil fertility. A tree with many uses, Leucaena leucocephala, has attracted much attention over the past decades. As per our literature review, no research has been conducted examining Leucaena leucocephala leaves for their fungal decomposition and their use as green manure. A study of the fungal colonization of Leucaena leucocephala leaves at various stages of decomposition was conducted to get an insight into which fungi play a critical role in the decomposition process. In total, fifty-two different species of fungi were isolated. There was an increase in the percentage of fungus occurrences as the leaves senesced and then finally decomposed. Almost all decomposition stages were characterized by a higher percentage occurrence of Deuteromycetes (75.47%) and by a lower rate of Ascomycetes (9.43%). A gradual increase of basidiomycetes such as unidentified sclerotia and Rhizoctonia solani was seen as the leaves senesced and finally decomposed. In the moist chamber, Didymium nigripes was the only Myxomycete isolated from completely decomposed leaves. In the present study, on average, there were more fungi in wet seasons than in the dry seasons.

1. Introduction

By 2050, we need to increase agricultural output by 70% to address international food security concerns and feed an exponentially growing global population. (https://sustainabledevelopment.un.org/post2015/transformingourworld, accessed on 15 March 2021). Various techniques currently in use to enhance crop yields, particularly the application of chemicals, has resulted in a wide range of negative impacts on both the environment and human health [1]. To mitigate the effects of chemical methods, mechanisms such as biological control [2] and biofertilization [3] are currently favoured. Biofertilization involves the cultivation of plants that produce green leaf manure, which aids in the betterment of the chemical, physical and biological properties of soil. In tropical countries such as India, the maintenance of good quality soil organic matter is a major concern and requires a steady input of organic residues to sustain soil fertility [4].
Leucaena leucocephala is a tree that has attracted a lot of attention in the past few decades due to its useful characteristics, such as a caloric value of 19.4 kJ/g and specific weight ranging from 0.50−0.59 kg/cm3. These properties make it conducive for paper and coal production [5,6]. It is utilised for making light structures and containers, as well as various types of fences and furniture such as tables, because it is easily machinable, porous to water-soluble preservatives and non-deformable after drying [5,7]. In addition, it is employed as a shade tree in a variety of plantations [7,8,9,10], as a mulch [11] and nitrogen fixer in the soil [12,13,14,15], as well as being used for regeneration of bare regions, slopes and pastures [9,16,17]. The leaves and seeds from Leucaena are also used as a food source in rural parts of Southeast Asia and Central America since they contain a high amount of fat (greater than 5.5%); particularly the fatty acids behenic, palmitic, stearic, linoleic, lignoceric and oleic acids [16,18]. It is also used as a beverage in place of coffee [12,19] and as a deworming agent [20,21]. A variety of brown, black and red pigments can be extracted from the bark, leaves and pods of Leucaena. On ingestion, it exhibits emmenagogic and abortive characteristics and is routinely used as a household medicine [22]. It is also beneficial for honeybees and exhibits defiance against dry climatic conditions in contrast to pastures and other forages that undergo browning and lose nutrients, thereby, ensuring availability of forage [23]. It is between 50% to 70% palatable and digestible [24] with a high nutritional range of 22−28% protein. It can, therefore, be used as a feed for both ruminants and non-ruminant livestock. Its high β-carotene and protein content along with its amino acid composition puts it on par with fodder obtained from alfalfa, fishmeal and soy meal [25]. It is composed of a few essential amino acids such as leucine, isoleucine, histidine and phenylalanine. The fodder is also a rich source of phosphorus, calcium and a number of other minerals [24,26,27,28], despite its lack of sodium [25,26]. A combination of total carbohydrates: 18.6%; starch: 1%; total oligosaccharides: 2.8%; reducing sugars: 4.2%; sucrose: 1.2% and raffinose: 0.6% has been reported by Kale in Akingbade [29].
The leaf surface habitat that harbours the wide range of both non-pathogenic and pathogenic microbes is called as the phylloplane [30,31]. Leaves become populated by an assortment of microorganisms from the moment they are formed and continue to sustain microbial populations through their lifetime. In the early stages of life, the leaf is possessed by several restricted or host specific parasites along with primary saprophytes [32]. These fungi obtain their nourishment either from the leaf itself or from the atmosphere. The fungi raid the readily decomposable sugars released from the surface of the leaf, faecal matter and honey dew from the leaf fauna—dead or decomposing parts of the leaf and healthy leaf tissue. The phylloplane fungi start disintegrating the cell walls and swiftly colonize the senescent leaf [33]. Very few fungi can be obtained from young leaves of the plants that are at seedling stage but as plants grow and leaves start to turn senescent the fungal population also exhibits exponential growth. The seed mycoflora is related to the epiphytic microflora of the above ground plant components from the time a plant grows from a seed [34]. The fact that treatment of seeds with fungicides suppresses epiphytic microbiota proves this.
Overall, two kinds of leaf colonizers are seen. The first are the airborne spores, called the “casual inhabitants”, that are there on the leaf surface simply by chance, while the second are the “resident inhabitants” that outnumber the casuals and are better adapted to the phylloplane [35,36]. According to Hudson [37], the pervasive fungi are those that are successful in establishing on the green leaves at an early stage and should be considered the primary saprophytes, while those with narrow host range are the restricted primary saprophytes.
According to available literature, no work exists till date to establish the fungal decomposition of leaves of Leucaena leucocephala and its use as green manure. The decomposition of organic matter of plant or animal origin is essentially a microbiological process in which fungi play an important role, so an assessment of the fungal colonization of Leucaena leucocephala leaves throughout its decomposition process was made to gain an understanding of the fungi that play a role in its decomposition. Considering that it is a perennial tree, this research may also be helpful in determining the impact of the environment on the dispersion of various fungi.

2. Materials and Methods

For the analysis of phylloplane of Leucaena leucocephala (Lam.) de Wit (var. K-8), the leaves were collected from Delhi state (28°4′ N and 77°2′ E) of India. Collections were made regularly at fortnightly intervals, throughout the year. Leaves were collected with a pair of sterilized forceps and scissors in sterile polythene bags and were brought to laboratory. Collections were made randomly from three different regions of trees (terminal, mid and lower regions of the tree).
Investigations were carried out on six categories of leaves:
Green leaves attached to the plant (GA).
Leaves beginning to yellow and senescence (YS).
Yellow and dry leaves just prior to abscission (YD) by spreading a plastic sheet under the tree and shaking it.
Yellow leaves immediately after abscission showing no sign of decomposition (YF).
Dried abscised leaves which were dark brown in colour and were partially decomposed, as indicated by disappearance of interveinal laminar region (PD).
Completely decomposed leaves which were black in colour due to almost total humification of laminar region. Care was taken to avoid collecting the soil associated with humified leaves (CD).
Phylloplane fungi were cultured in moist chambers for identification. For preparing moist chambers 3–4 filter papers were kept in Petri plates and were sterilized at 15 lbs. for fifteen minutes. These were moistened with sterile distilled water. Five leaves were placed in each plate under sterile conditions. Three replicates were maintained for each treatment. The moist chambers with leaves were incubated at 26 ± 1 °C for five days [38].
For quantitative estimation of fungi, the dilution plate method [39,40] was employed. One gram of leaves collected randomly at the six different stages were taken separately in 250 mL flasks containing 100 mL of sterile distilled water. The flasks were mechanically shaken for 1 h. The water after washing was decanted and serial dilutions were prepared and used for plating. The plating of 1:1000 dilution was found to be appropriate for colony counting. The plating was done using PDA and Czapek’s Dox Agar medium to get the maximum number of fungi. The entire procedure was done under sterile conditions. Colonies appearing in the culture plates, whether from spores or mycelia, were counted. Microscopic studies were done after 5 days of incubation.
For quantitative estimations, the number of fungal propagules were expressed as the number of colonies/g weight of leaves. It was calculated as
A v e r a g e   n u m b e r   o f   c o l o n i e s   p e r   p e t r i p l a t e A m o u n t   o f   A l i q u o t   u s e d × D i l u t i o n   o f   t h e   a l i q u o t D r y   w e i g h t   o f   t h e   l e a v e s
For qualitative estimations, percent frequency of occurrence of various fungi and relative frequency occurrence of the fungal groups were calculated at different stages of leaf decomposition as well as during different months of the year.
Percentage of frequency was calculated by using the formula given by Tresner et al. (1954).
%   Frequency = N u m b e r   o f   s a m p l e s   w i t h   f u n g u s   s p e c i e s T o t a l   n u m b e r   o f   s a m p l e s × 100
Relative percent frequency of occurrence of various groups of fungi viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes in different stages of decomposition was calculated as
Relative   %   Frequency = T o t a l   p e r c e n t   f r e q u e n c y   o f   o c c u r r e n c e   o f   a   g r o u p T o t a l   p e r c e n t   f r e q u e n c y   o f   o c c u r r e n c e   o f   a l l   f u n g a l   g r o u p s × 100 .

3. Results

In all, fifty-two different fungi were isolated using various techniques. Direct observation of the leaves showed few conidia of Alternaria alternata, chains of cells of Aureobasidium pullulans and acervuli of Colletotrichum. Along with these, mycelial growth could also be seen. Leaves are known to be colonized since the first unfolding of the leaves. However, only a few fungi are present at this stage. As the plant ages, there is an increase in the number of fungal colonies per gram of the leaves. Table 1, Table 2, Table 3, Table 4, Table 5 and Table 6 and Figure 1, Figure 2, Figure 3, Figure 4, Figure 5 and Figure 6 give the complete sequential changes in fungal population during different stages of plant growth as well as in different months of the year.
Average percent frequency of occurrence of various fungi showed a gradual increase as the leaves senesced and finally decomposed completely.
The saprophytic population on mature green leaves (GA) (Table 1) was comprised mostly of Phaeoramularia graminicola, Fusarium lateritium, Aureobasidium pullulans, Cladosporium cladosporioides and Arthrinium cuspidatum. These were followed in frequency of occurrence by Aspergillus niger, Alternaria alternata and Penicillium chrysogenum. There was also an occasional or rare appearance of a few species of Aspergillus, Candida albicans, Chaetomium globosum, Pestalotia monorhincha, Epicoccum nigrum, Nigrospora sphaerica, Drechslera tetramera and Myrothecium roridum. As the leaves senesced (YS) (Table 2) the percent frequency of occurrence of Aureobasidium pullulans, Cladosporium cladosporioides, Aspergillus niger, A. flavus, Alternaria alernata, Drechslera tetramera, Aspergillus nidulans, Gloeosporium sp. and Phoma hibernica increased, whereas Colletotrichum falcatum, Gliocladium atrum, Cephalosporium acremonium, Mucor hiemalis and Monilia geophila joined the microbial community at the later stages. During drying of the leaves before leaf fall (YD) (Table 3) there was not much colonization of the leaves by new fungi, but some of the pre-existing saprophytes like Chaetomium globosum, Pestalotia monorhincha, Fusarium lateritium and Epicoccum nigrum disappeared from the community and Gloeosporium sp. was the most frequently occurring fungus at this stage. The freshly fallen Leucaena leaves (YF) (Table 4) appeared to be less extensively colonized by commonly accepted phylloplane fungi than when they were attached to the tree. Some of the existing fungi still persisted there and showed an increase in their percentage frequency of occurrence viz. A. niger, Fusarium equiseti, whereas a few like Alternaria alternata, Cladosporium cladosporioides, Phaeoramularia graminicola and Aureobasidium pullulans showed a decline in their percent frequency of occurrence, which may be due to interactions among themselves.
After the leaves started decomposing (PD) (Table 5), there was a slight change in the existing mycoflora of the leaves; Absidia repens, F. lateritium, Myrothecium roridum and Cladosporium cladosporioides dominated the fungal flora. Other existing saprophytes like Colletotrichum falcatum, Mucor hiemalis and Trichoderma viride showed an increased percent frequency of occurrence. Alternaria alternata, Aspergillus nidulans and Gloeosporium sp., however, occurred only occasionally.
After the complete decomposition of the leaves (CD) (Table 6), the most common fungi included Cladosporium cladosporioides, Colletotrichum falcatum, species of Fusarium, Myrothecium roridum and Phoma hibernica. The characteristic feature was the appearance of myxomycete Didymium nigripes. This fungus did not appear at any other stage of decomposition. The percent frequency of occurrence of Mucor hiemalis increased greatly and there was occasional appearance of Memnoniella echinate and Stachybotrys atra which could not be isolated from any other stage of decomposition. Species of Penicillium were higher in green leaves, and as the leaves decomposed their percent frequency declined.
The fungi isolated were classified into various fungal groups viz., Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes. The relative frequency of occurrence of various groups are shown in Figure 1, Figure 2, Figure 3, Figure 4, Figure 5 and Figure 6. It is evident from the figures that the relative percent frequency of occurrence of Deuteromycetes (75.47%) was highest at almost all the decomposition stages and that of Ascomycetes (9.43%) was the lowest. There was gradual increase in the occurrence of Deuteromycetes from when the leaves began to senesce up to their fall. But as leaves underwent decomposition their percent frequency of occurrence showed a decline. Corresponding to the decline in Deuteromycetes, the relative frequency of Zygomycetes increased sharply after the leaves had fallen and decomposed partially. However, after the complete decomposition of leaves, they showed a slight decrease from the partially decomposed stage. The relative percent frequency of occurrence of Ascomycetes was highest when the leaves had senesced but were still attached to the tree. Immediately after the leaves fell there was a sudden decrease in the value, which was followed by a sharp increase. Basidiomycetes, which were represented by sterile mycelium, unidentified sclerotia form and Rhizoctonia solani, showed a gradual increase as the leaves senesced and finally decomposed completely. Myxomycetes were represented by Didymium nigripes, which was isolated only from the completely decomposed leaves in the moist chamber.
Seasonal Variation of the Various Fungi Colonising Different Decomposition Stages is shown in Table 7 and Table 8.
Mainly three seasons affect the microfungal population on leaves i.e., rainy, winter and summer. The percent frequency of occurrence of various fungi in different seasons at different decomposition stages were calculated and then grouped into five categories of frequency:
1.0–20%Rare
2.21–40%Occasional
3.41–60%Frequent
4.61–80%Common
5.81–100%Very common
Green leaves which were attached on the tree (GA)—As observed in Table 7 and Table 8, a few species like Alternaria alternata, Penicillium chrysogenum, Cladosporium cladosporioides occurred throughout the year but with varying frequencies. Aureobasidium pullulans and Phoma hibernica were most common in the rainy season, declining in frequency during the winters and being completely absent in the summers. Penicillium chrysogenum occurred rarely in summers.
In winters the most frequent fungus was Cladosporium cladosporioide, Candida albicans, Cicinella muscae and Chaetomium globosum occurred rarely and only during the rainy season. Nigrosopora sphaerica and Arthrinium cuspidatum were strictly seasonal and could be isolated only during the winters.
Yellow leaves which were still to the tree (YA)—At this decomposition stage, Phaeoramularia graminicola, which represented the most frequent fungus of green mature leaves during the rainy season, showed overall decline in its percent frequency of occurrence. Aureobasidium pullulans, Alternaria alternate, a species of Aspergillus, Penicillium chrysogenum and Phoma hibernica represented the most frequent flora during rainy and winter seasons. Myrothecium roridum, Nigrospora sphaerica and Epicoccum nigrum represented the strictly winter flora. Cladosporium cladosporioides was dominant in winters as well as at the beginning of summers, and Epicoccum showed a decline in its percent frequency of occurrence. Aspergillus nidulans was present in the rainy season, and only once in the month of May.
Yellow Senescent leaves prior to their fall on the ground (YD)—Phaeoramularia graminicola was the common fungus in this stage of decomposition also. Aureobasidium pullulans, Alternaria alternata, Cladosporium cladosporioides and Penicillium chrysogenum frequently occurred in all the seasons of the year. Nigrospora and Arthrinium represented the dominant flora of winter only. The frequency of Aspergillus nidulans increased during summers. Colletotrichum falcatum could be isolated only occasionally or rarely in the rainy season.
Yellow Leaves which were recently fallen on the ground (YF)—This decompositional stage of leaves shows the predominance of Aspergillus luchuensis, Penicillium chrysogenum and Phaeoramularia during the rainy season. Gloeosporium sp., which represented the most frequent fungus in this stage, could be isolated throughout the year. Some of the dominant fungi of winters like Arthrinium cuspidatum and Nigrospora sphaerica, however, occurred very rarely in this decomposition stage. Cladosporium cladosporioides and Aureobasidium pullulans were the dominant fungi in winters.
Partially Decomposed Leaves (PD)—Colletotrichum falcatum was the dominant fungus at this stage in the rainy season, with a few species of Aspergillus. Cladosporium cladosporioides, along with Aureobasidium pullulans and Gloesporium sp., were among the most frequent fungi in winter flora. Alternaria alternata showed decreased percent frequency of occurrence throughout. In summers, Myrothecium roridum and Absidia repens were among the most dominant fungi, with Monilia geophila occurring rarely.
Completely Decomposed Leaves (CD)—Colletotrichum falcatum, Didymium nigripes and Myrothecium roridum were the most frequent fungi of rainy season. In winters Cladosporium cladosporioides was the dominant fungus at this stage also. Mucor hiemalis and Rhizopus stolonifer showed greatly enhanced percent frequency of occurrence, the maximum of which could be isolated in winters. Aspergillus pullulans was not isolated from leaves of this stage at all. Alternaria alternata could also be isolated only rarely. However, occasional occurrence of Memnoniella echinata and Stachybotrys atra could be seen during the winters. The summer fungal flora showed the predominance of Cladosporium cladosporioides, Myrothecium roridium, Phoma hibernica and Colletotrichum falcatum.

4. Discussion

During the present study, the fungi were isolated from leaves at six different decomposition stages in different seasons of the year. It is evident from the data that by the time the leaves of Leucaena reach the surface litter, they are substantially colonized by a variety of parasitic and saprophytic fungi. This contrasts with Hering’s [41] conclusion that the fungi dominant on fallen oakwood litter were not present to any great extent before leaf fall. However, leaves of Quercus rotundifolia Lam [42], Pinus sylvestris [43,44], Populus tremuloides [45], Ilex aquifolium [46], Sesamum orientale, Gossypium hirsutum [40] and Mangifera indica [47] have colonization of leaves by fungi before their fall.
According to Hudson’s scheme [37], the primary saprophytic colonizers are represented by Ascomycetes and Deuteromycetes, of which Cladosporium herbarum, Alternaria alternate, Epicoccum nigrum, Aureobasidium pullulans and Botrytis cinerea are common components of successions. These are present on the phylloplane of living leaves as spores, which become vegetatively active only at leaf senescence. Most of these common fungi were isolated from leaves of Leucaena also. However, in the present investigation, colonization of leaves by these fungi occurred at very early stage, as was also reported by Dickinson [48], Lindsey and Pugh [49], Mishra and Dickinson [46] Promputtha, et al. [50] and Wildman and Parkinson [45]. Therefore, commencement of pathogenic activity did not coincide with leaf senescence. Initially, at the seedling stage, there were very few fungal propagules on the leaves. As the plant aged, the number of fungal colonies per gram of leaves increased, which may be due to prolonged exposure of the leaves to the air spores, and it is also believed that ageing leaves produce a greater number of exudates which enhance fungal colonization. As the leaves senesced, there was a gradual increase in internal colonization of the leaves, indicating a slow penetration of the fungi into leaves with time. Heavy colonization of senescing leaves has also been reported by several other works [40,46,49,50,51,52].
After the leaves fell on the ground and their decomposition was initiated, the primary saprophytes on leaves were joined by new colonizers and a few pre-existing fungi showed increased percentage frequency of occurrence. Some of the new colonists included Absidia repens, Bipolaris sp. Stachybotrys atra, Memnoniella echinata, Mucor hiemalis and Didymium nigripes, which were present on litter. The pre-existing species like Pestalotia monorhincha, Epicoccum nigrum and Fusarium lateritium could not be isolated from the litter.
The persistence of primary saprophytes during the initial decay period has been attributed to various factors. The period of the parasitic phase, which enables them to penetrate and establish in freshly decaying tissue, is very short. According to Hogg [53] and Visser and Parkinson [54] most of the primary saprophytes have a high rate of sporulation, and due to their ability to survive under drought conditions, they persist for a longer time. Higher sporulation in the fungi was probably due to the availability of a large amount of soluble nutrients.
Extensive work has been done on various aspects of the microbial ecology of leaf and litter in relation to the degradation and fertility of soil [4,50,55,56,57]. However, much is not known about the decomposition potentials of the fungal colonizers. Sharma and Mukerji [58,59] recognized four different types of fungal colonization patterns on leaves of Sesamum orientale and Gossypium hirsutum. Differences were determined chiefly by the developmental stage of the organ on which the organism occurred, the substrate it provided, the capacity of the fungus to utilize the available substrates, and the potential of the fungus to degrade the organ following senescence.
In the current study, Alternaria alternata, Cladosporium cladosporioides, different species of Fusarium, Aspergillus, Penicillium chrysogenum, Trichoderma viride and Phoma hibernica appeared to have no restricted distribution. They occurred significantly during the living as well as the senescent and decaying phases of the leaves. Although Penicilli, Aspergilli and Trichoderma viride occurred quite significantly in dilution plates, these were absent or rare in moist chambers. Dickinson [60] and Webster [61] classified Aspergilli and Penicilli as casual inhabitants which play negligible roles in decomposition. Sharma et al. [40] could isolate Trichoderma viride very rarely, and they grouped it under non-decomposers, but Hering [41] showed that Penicillium sp. And Trichoderma viride, though isolated only in dilution plates, did have a role in decomposition, and are important cellulose degrading fungi.
The completely decomposed leaves incubated in moist chambers were peculiar in that they were exclusively colonized by the sporulating stages of Didymium nigripes. This agrees with earlier reports on cotton leaves and on leaves and stems of Gossypium and Sesamum [62]. The plating of surface washings of decaying organs exhibited a high number of imperfect fungi, especially those which have been considered decomposers [58].
Another interesting observation was the frequent isolation of Mucorales from the surface washing of decaying leaves. The frequency of Mucorales significantly increased on highly decomposed leaves. Similar results were obtained by Sharma and Mukerji [62]. However, Sharma et al. [40] has not mentioned the position of yeasts like fungi viz. Aureobasidium pullulans in a successional pattern. Phaeoramularia graminicola was also isolated very frequently in the present study on attached leaves. With leaf fall, there was a decline in frequency of occurrence of Aureobasidium pullulans. This decline in yeast and Aureobasidium frequency may have been due to the dryness of the fallen leaves. Ruscoe [44] used a direct observation technique and showed that Aureobasidium colonized the phylloplane of young Nothofagus leaves. This fungus formed vigorously, growing colonies whose development increased with increasing leaf age and then declined at leaf fall. However, autolysis of fungal hyphae was not reported by Wildman and Parkinson [45].
Garrett [63] proposed a generalized fungal sequence on plant material within or on the soil. The sequence is as follows: weak parasites → primary saprophytic sugar fungi → cellulose decomposers → secondary sugar fungi → lignin decomposers + associated fungi.
The colonization pattern of Leucaena leaves at different decomposition stages followed the general trend. The green mature leaves were colonized predominantly by Deuteromycetes, along with the rare appearance of other groups of fungi. As the leaves senesced and decomposed, the soil Ascomycetes (which are known to be important cellulose degraders) increased in their percent frequency of occurrence. Litter in its final stages of decomposition showed enhanced growth of Mucorales and Basidiomycetes. The Myxomycetes isolated in the present study and reported earlier [62,64] from the completely decomposed leaf litter, however, represent the secondary saprophytes of the Hudson’s scheme [37].
Since Leucaena is an evergreen tree, all kinds of attached and fallen leaves were present at the same time. It may be that some fungi are found in all stages of leaf decomposition, suggesting that there is a two-way relationship between the mycoflora of living and dead leaves. These will affect successful establishment, growth, reproduction and survival of both phylloplane inhabitants and litter decomposers.
Environmental factors play a very important role in quantitative and qualitative distribution of the fungi on the leaves. In the present study, average number of fungi were more in wet season than in the dry season. The greater number of fungal taxa recorded in wet season is like the observations of Almaguer, et al. [65,66], Jothish & Nayar [67] and Stennett & Beggs [68]. This may be due to high relative humidity, moderate temperature and lower sunshine duration. According to Diem [69], in the rainy season the cuticle is constantly wet, which is suitable for the growth of fungi. Irrespective of plant age, several fungi occurred only during a specific period of investigation. For example, Phaeoramularia graminicola and Didymium nigripes were strictly isolated in the rainy season. Arthrinium cuspidatum, Nigrospora sphaerica, Epicoccum nigrum and Pestalotia monorhincha were observed only in the winters. It is thus evident that microbial activity depends on the micro and macro environmental conditions and the substrate characteristics. Studies on the associated fungi of the same plant grown in different localities, therefore, is of great interest.

5. Conclusions

Average percent frequency of occurrence of various fungi gradually increased as the leaves senesced and finally decomposed completely. The mature green leaves were colonized by Chaetomium golobosum, Pestalotia monorhincha, Fusarium lateritium and Epicoccum nigrum along with Phaeoramularia graminicola, Aureobasidium pullulans, Alternaria alternate and Cladosporium sp., which were very frequent. As the leaves senesced, the percentage frequency of occurrence of a few pre-existing fungi increased, whereas Colletotrichum falcatum, Gliocladium atrum, Cephalosporium acremonium, Mucor hiemalis and Monilia geophila later joined the microbial community. Upon drying the leaves before leaf fall there was not much colonization of the leaves by new fungi, but some of the pre-existing fungi like Chaetomium globosum, Pestalotia monorhinch, F. lateritium and Epicoccum nigrum disappeared from the community and Gloeosporium appeared, which was the most frequently occurring fungus at this stage. The freshly fallen Leucaena leaves appeared to be less extensively colonized by commonly accepted phylloplane fungi than when they were on the tree. After the leaves started decomposing there was slight change in the existing mycoflora of the leaves. Absidia, Fusarium lateritium, Myrothecium roridum and Cladosporium sp. dominated the fungal flora. After complete decomposition of the leaves, the characteristic feature was the appearance of Myxomycetes Didymium nigripes. Memnoniella echinata and Stachybotrys atra could also be isolated only from this stage of decomposition. In the present study, on average, the number of fungi was greater in the wet season than in the dry season.

Author Contributions

S.G. and J.K. conceptualized and conceived the idea. S.G., J.K., P.C., M.A.P., N.B., P.M., R.K., C.D.R., S.K. compiled and formulated the manuscript. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Acknowledgments

Authors are thankful to their respective institutions for facilities and support. The authors acknowledge the contributions of Nellie Laisram and Mallika Kalia in the preparation of the manuscript.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at GA Stage.
Figure 1. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at GA Stage.
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Figure 2. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at YS Stage.
Figure 2. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at YS Stage.
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Figure 3. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at YD Stage.
Figure 3. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at YD Stage.
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Figure 4. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at YF Stage.
Figure 4. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at YF Stage.
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Figure 5. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at PD Stage.
Figure 5. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at PD Stage.
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Figure 6. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at CD Stage.
Figure 6. Relative % frequency of occurrence of various groups of fungi on leaves of Leucaena leucocephala (viz. Deuteromycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Myxomycetes) at CD Stage.
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Table 1. Percentage frequency of occurrence of various fungi on Green Attached Leaves (GA) of Leucaena leucocephala.
Table 1. Percentage frequency of occurrence of various fungi on Green Attached Leaves (GA) of Leucaena leucocephala.
Months of the Year
FungiJulyAug.Sept.Oct.Nov.Dec.Jan.Feb.Mar.Apr.MayJune
DEUTEROMYCETES
Alternaria alternata90805030406050602010030
Arthrinium cuspidatum 60
Aspergillus flavus3020501010 40
A. fumigatus20 10 50
A. humicola 20
A. luchuensis3020 30
A. niger8080505010 508020 5020
A. sulphureus2030 1030 20
Aureobasidium pullulans90 100409010070301020
Candida albicans10
Cladosporium cladosporioides 1080100100806060
Curvularia lunata204080 20
C. pallescens 20
Drechslera tetramera20101020
Drechslera hawaiiensis 10
Epicoccum nigrum 10203050
Fusarium lateritium 100
F. oxysporum 20501010 30 60
Glaeosporium sp. 105020
Myrothecium roridum 10 20 30
Nigrospora sphaerica 601040
Penicillium chrysogenum30909080 2010
P. citrinum 20
P. funiculosum 40 10 20
Pestalotia monorhincha 30 30
Phaeoramularia graminicola 80100
Phoma hibernica3050502040
Trichoderma viride 1010
ZYGOMYCETES
Choanephora cucurbitarum10 50
Circinella muscae 10
Rhizopus stolonifer 10 10 20 2020
ASCOMYCETES
Chaetomium globosum 10
Aspergillus nidulans5030 202020 30
BASIDIOMYCETES OR MYCELIA STERILIA
Sterile mycelium102030 70 6030 50 30
Unidentified Sclerotial form 10
Table 2. Percentage frequency of occurrence of various fungi on yellow attached Leaves (YS). Leucaena leucocephala.
Table 2. Percentage frequency of occurrence of various fungi on yellow attached Leaves (YS). Leucaena leucocephala.
Months of the Year
FungiJulyAug.Sept.Oct.Nov.Dec.Jan.Feb.Mar.Apr.MayJune
DEUTEROMYCETES
Alternaria alternata608020 5060 707060
Arthrinium cuspidatum 20
Aspergillus flavus905020 10
A. fumigatus50 20
A. luchuensis10030 20
A. niger90808010 302050
A. sulphureus 20
A. terreus10
Aureobasidium pullulans 5080 10060
Cephalosporium reseogriseum 10
Cladosporium cladosporioides 60 1001001009010080
Colletotrichum falcatum 4020
Curvularia lunata30
Drechslera tetramera3060
Epicoccum nigrum 20 20
Fusarium oxysporum60 40 20 10
F. semitectum10 20 30
Gloeosporium sp. 40805020
Gliocladium atrum 40
Myrothecium roridum 50
Nigrospora sphaerica 10080806020
Penicillium chruysogenum30906020
P. citrinum 20 10
P. funiculosum10 10
Phaeoramularia graminicola 8020
Phoma hibernica 6060 10020203020
Trichoderma viride101010
ZYGOMYCETES
Circinella muscae20
Mucor hiemalis10
Rhizopus stolonifer2010
ASCOMYCETES
Aspergillus nidulans6030 50
BASIDIOMYCETES OR MYCELIAL STERILIA
Sterile mycelium606040 2020102020
Unidentified Sclerotial form 50
Table 3. Percentage frequency of occurrence of various fungi on leaves before they fall on ground (YD).
Table 3. Percentage frequency of occurrence of various fungi on leaves before they fall on ground (YD).
Months of the Year
FungiJulyAug.Sept.Oct.Nov.Dec.Jan.Feb.Mar.Apr.MayJune
DEUTEROMYCETES
Alternaria alternata105010 1010010090100902010
Arthrinium cuspidatum 3020
Aspergillus flavus100404090 302030‘
A. fumigatus70
A. luchuensis70 20 10
A. niger100805080 303040
A. sulphureus 30
A. terreus10
Aureobasidium pullulans2010 1010060 60100
Cladosporium cladosporioides 50100100100809080 10
Colletotrichum falcatum 20 40
Curvularia lunata20301020 30
Cylindrocarpon sp.10
Drechslera tetramera 10
Epicoccum nigrum 20 80
Fusarium oxysporum6010502010
Gloeosporium sp. 100
Monilia geophila 10 1020
Myrothecium roridum 10 20
Nigrospora sphaerica 10802060 10
Penicillium chrysogenum30809070 302020
P. funiculosum1040 10 10
Phaeoramularia graminicola 6090
Phoma hibernica 10 109020
Trichoderma viride10 10
ZYGOMYCETES
Choanephora cucurbitarum 20
Rhizopus stolonifer10 10 10
ASCOMYCETES
Aspergillus nidulans303010 2020205010
BASIDIOMYCETES OR MYCELIA STERILIA
Sterile mycelium 504010 30 50
Unidentified sclerotial form 30
Table 4. Percentage frequency of occurrence of various fungi on yellow-Fallen leaves (YF).
Table 4. Percentage frequency of occurrence of various fungi on yellow-Fallen leaves (YF).
Months of the Year
FungiJulyAug.Sept.Oct.Nov.Dec.Jan.Feb.Mar.Apr.MayJune
DEUTEROMYCETES
Alternaria alternate4050 1020 4030
Arthrinium cuspidatum 10
Aspergillus flavus6040 50 101030
A. fumigatus70 40 70
A. humicola 10
A. luchuensis100 20
A. niger100806070 205060
A. sulphureus10 10 20 50
A. terreus 10
Aureobasidium pullulans1010 1010010090701010
Choanephora cucurbitarum 20
Cladosporium cladosporioides 201001001009060102030
Colletotrichum falcatum 20602040 50 20
Curvularia lunata103040 10
C. pallescens 10
Cylindrocarpon sp.4010
Drechslera tetramera30
Epicoccum nigrum 20
Fusarium equiseti60 80
F. lateritium 9010
F. oxysporum 1020 10
F. semitectum 1020 80
Gloesporium sp. 100 10060607080
Mucor hiemalis 20 102010
Myrotehcium roridum 10 1002020
Nigrospora sphaerica 20
Penicillium chrysogenum108010040
P. funiculosum1040 10
P. nigricans20
Phaeoramularia grominicola 6080
Phoma hibernica 1010 20 30
Trichoderma viride 1020 10
ZYGOMYCETES
Choanephora cucurbitarum 20
Rhizopus stolonifer10 50 10 10
ASCOMYCETES
Aspergillus nidulans2030
BASIDIOMYCETES OR MYCELIA STERILIA
Sterile mycelium4040 10 20 605050
Rhizoctonia solani 10 4010
Table 5. Percentage frequency of occurrence of various fungi on partially decomposed leaves (PD).
Table 5. Percentage frequency of occurrence of various fungi on partially decomposed leaves (PD).
Months of the Year
FungiJulyAug.Sept.Oct.Nov.Dec.Jan.Feb.Mar.Apr.MayJune
DEUTEROMYCETES
Alternaria alternata 10 1020 70
Aspergillus flavus100901080 206050
A. fumigatus9020 100
A. luchuensis10010 20 90
A. niger1001004010020 2030 8090100
A. sulphureus 30
A. terreus 20
Aureobasidium pullulans 8040 50 10
Cladosporium cladosporioides 8010010080808040
Colletotrichum falcatum10080 10 30
Curvularia lunata 10 20
Cylindrocarpon sp. 20
Drechslera tetramera10
Fusarium lateritium 10080
F. oxysporum30 2080
F. semitectum 202020
Gloeosporium sp. 1020 60
Monilia geophila 60 20
Myrothecium roridum 90
Nigrospora sphaerica 10
P. chrysogenum 104010 40
Penicillium citrinum 40
P. funiculosum 10
Phoma hibernica 1060
Trichoderma viride10 40 20
ZYGOMYCETES
Absidia repens 100
Choanephora cucurbitarum102010
Mucor hiemalis 10 100 60604010405010
Rhizopus stolonifer20 10 2010
ASCOMYCETES
Aspergillus nidulans2010 8020
BASIDIOMYCETES OR MYCELIA STERILIA
Sterile mycelium 20 20
Table 6. Percentage frequency of occurrence of various fungi on completely decomposed leaves (CD).
Table 6. Percentage frequency of occurrence of various fungi on completely decomposed leaves (CD).
Months of the Year
FungiJulyAug.Sept.Oct.Nov.Dec.Jan.Feb.Mar.Apr.MayJune
DEUTEROMYCETES
Alternaria alternata 40 20
Aspergillus flavus9090509080 1001006070
A. fumigatus4010 10 100
A. luchuensis20203030 10 80
A. niger90100100100 504010040100
A. parasiticus 10
A. sulphureus 90 40 100
A. terreus 1040 20
Bipolaris sp. 10
Cladosporium cladosporioides 1008010080100100
Colletotrichum falcatum70 100 90 10080
Fusarium lateritium 100
F. oxysporum90 40 50
F. semitectum 50 20 60 5070 40
Memnoniella echinata 40
Myrothecium roridum100
Penicillium chrysogenum 50 90
P. citrinum 30
Phaeoramularia graminicola 10
Phoma hibernica 90
Stachybotrys atra 1010
Trichoderma virde 4030 10
ZYGOMYCETES
Absidia repens 40
Choanephora cucurbitarum 3010
Mucor hiemalis2030 100 40 50405010
Rhizopus stolonifer1040 30 60 60 10
ASCOMYCETES
Aspergillus nidulans20 50
BASIDIOMYCETES OR MYCELIA STERILIA
Sterile mycelium1090202030 80
MYXOMYCETES
Didymium nigripes100
Table 7. Average percentage frequency of occurrence of various fungi at different decomposition stages of the leaves in different seasons.
Table 7. Average percentage frequency of occurrence of various fungi at different decomposition stages of the leaves in different seasons.
FungiGreen Attached (GA)Yellow Attached (YA)Yellow before Fall (YF)Recently Fallen (RF)Partially Decomposed (PD)Completely Decomposed (CD)
RWSRWSRWSRWSRWSRWS
Abisidia repens100
Alternaria alternata7348505355672380554515351015704020
Arthrinium cuspidatum60202510
Aspergillus flavus3310405325609027335027678043778583
A. fumigatus15505020707027551002555
A. humicola2010
A. luchuensis25306520702010100205590233045
A. niger704830831033778033807043804390977570
A. parasiticus10
A. sulphureus2520202030101035309070
A. terreus1010102023
Aureobasidium pullulans956615658015581001074106030
Basidiomycete (Sclerotial)105030
Bipolaris sp.10
Candida albicans10
Cephalosporium roseogriseum10
Chaetomium globosum10
Choanephora cucurbitarum302010201320
Circinella muscae1020
Cladosporium cladosporioides108460609890509445209020886092100
Colletotrichum falcatum3020404030359010308590
Curvularia lunata47203030203027101020
C. pallescens2010
Cylindrocarpon Sp.102520
Didymium nigripes100
Drechslera tetramera1320451030303010
D. hawaiiensis10
Aspergillus nidulans4020204550352050’2515502050
Epiococcum282020208020
Fusarium equiseti6080
F. lateritium1005090100
F. oxysporum3510455020104015151030506550
F. semitectum10203015802020504053
Gliocladium atrum40
Gloeosporium Sp.3020405010010073801560
Memnoniella echinata40
Monilia geophila1040
Mucor hiemalis102033106540256333
Myrothecium roridum10203050102010479010090
Nigrospora sphaerica03780201053102010
Penicillium chrysogenum70801560206740206340251050
P. citrinum102020104030
P. funiculosum40201010251010251010
P. nigricans20
Pestalotia monorhincha3030
Phaeoramularia graminicola9050757010
Phoma hibernica653060432010401020303590
Rhizoctonia solani20
Rhizopus stolonifer10152010101010103010201015254535
Stachybotrys atra10
Sterile mycelium20534053182033305040205320202780
Trichoderma viride1010101020101040203010
Table 8. Seasonal variation of various fungi at different decomposition stages of the leaves.
Table 8. Seasonal variation of various fungi at different decomposition stages of the leaves.
Different Decomposition StagesSeasonVery CommonCommonFrequentOccasionalRare
Green Leaves Which Were Attached on the Tree (GA)Rainy Season Aureobasidium pullulans, Phaeoramularia graminicolaAspergillus niger, Alternaria alternate, Penicillium chrysogenum, Phoma hibernicaCurvularia lunataAspergillus flavus, A. luchuensis, A. sulphureus, Aspergillus nidulans, Penicillium funiculosum, Fusarium oxysporum, Gloeosporium sp., Pestalotia monorhincha.Candida albicans, Chaetomium globosum, Circinella muscae, Cladosporium sp., Drechslera tetramera, D. hawiiensis, Myrothecium roridum, Rhizopus stolonifer, sterile mycelium.
Winter SeasonCladosporium cladosporioidesAureobasidium pullulans, P. chrysogenumA. niger, Alternaria alternate, Arthrinium cuspidatum, Sterile mycelium.Epicoccum nigrum, Nigrospora sphaerica, Pestalotia monorhinca, Phoma hibernica.Aspergillus flavus, Curvularia lunata, C. pallescens, Drechslera tetramera, Aspergillus nidulans, F. oxysporum, Gloeosporium sp. Myrothecium roridum, P. citrinum, Rhizopus stolonifer, Trichoderna viride.
Summer SeasonFusarium lateritiumNoneAspergillus fumigatus, Alternaria alternate, Cladosporium cladosporioide, Fusarium oxysporumAspergillus flavus, A. luchuensis, A. niger, sterile mycelium.Aspergillus humicola, A. sulphureus, Aspergillus nidulans, Penicillium chrysogenum, P. citrinum, Rhizopus stolonifer, unidentified Basidomycetes
Yellow Leaves which were still attached on the tree (YA)Rainy SeasonAspergillus nigerAspeurgillus luchensis, Aureobasidium pullulansAspergillus flavus, A. fumigatus, Alternaria alternate, Cladosporium cladosporioides, Drechslera tetramera, Aspergillus nidulans, Fusarium oxysporum Penicillium chrysogenum, Phaeoramularia graminicola, Phoma hibernica, sterile mycelium, unidentified BasidiomyceteCurvularia lunata, Gloesporium sp., Gliocladium atrumAspergillus terreus, Circinella muscae, Fusarium semitectum, Mucor hiemalis, Penicillium funiculosum, Rhizopus stolonifer, Trichoderma viride
Winter SeasonCladosporium cladosporioidesAureobasidium pullulans, Nigrospora sphaericaAlternaria alternate, Gloeosporium sp., Myrothecium roridum, Phoma hibernicaCollectotrichum falcatumAspergillus niger, Cephalosporium roseogriseum, Epicoccum nigrum, Fusarium oxysporum, F. semitectum, Penicillium chrysogenum, P. citrinum, sterile mycelium.
Summer SeasonCladsoporium cladosporioidesAlternaria alternateAspergillus nidulansAspergillus flavus, A. niger, Fusarium semitectumAspergillus fumigatus, A. luchensis, A. sulphureus, Arthrinium cuspidatum, Epicoccum nigrum, Fusarium oxysporum Nigrospora sphaerica, penicillium citrinum, P. funiculosum Rehizopus nigricans, sterile mycelium.
Yellow Senescent Leaves prior to their fall on the ground (YD)Rainy SeasonGloeosporium sp.Aspergillus fumigatus, A. luchuensis, A. niger, Penicillium chrysogenum, Phaeoramularia graminicolaAspergillus flavus, Cladosporium cladosporioidesAlternaria alternate, Curvularia lunata, Aspergillus nidulans, Fusarium oxysporum, Penicillium funiculosum, sterile mycelium.Aspergillus terreus, Aureobasidium pullulans, Choanephora cucurbitarun, Colletotrichum falcatum, Cylindrocarpon sp., Drechslera tetramera, Myrothecium roridum, Nigrospore sphaerica, Phoma hibernica, Rhizopus stolonifer, Trichoderma viride.
Winter SeasonAspergillus flavus, Cladosporium cladosporioidesAspergillus niger, Alternaria alternataAureobasidium pullulans, Nigrospora sphaericaArthrinium cuspidatum, Penicillium chysogenum, Phoma hibernica, Colletotrichum falcatum, sterile myceliumAspergillus luchuensis, Curvularia lunata, Aspergillus nidulans, Epicoccum nigrum, Fusarium oxysporum, Monilia geophila, Myrothecium roridum, Penicillium funiculosum,
Summer SeasonAirepnasodoi pullulansEpicoccum nigrumAlternaria alternate, Cladosporium cladosporioides, Aspergillus nidulans, sterile myceliumAspergillus flavus, A. niger, A. sulphureus, Curvularia lunata, Drechslera tetramera, unidentified Basidiomcete.Aspergillus luchuensis, Choanephora cucurbitarum, Nigrospora sphaerica, Penicillium chrysogenum, P. funiculosum, Rhizopus stolonifer.
Yellow Leaves which were recently fallen on the ground (YF)Rainy SeasonAspergillus luchuensis, Gloeosporium sp.Aspergillus fumigatus, A. niger, Phaeoramularia graminicola, Penicillium charysogenumAlternaria alternate, Fusarium equisetiAspergillus flavus, Colletotrichum falcatum, Curvularia lunata, Cylindrocarpon sp., Drechslera tetramera, Emericellan nidulans, Penicillium funiculosum, sterile mycelium.Aspergillus sulphureus, Choanephora cucurbitarum, Aureobasidium pullulans, Cladosporium cladosporioides, Fusarium oxysporum, Myrothecium roridum, Penicillium nigricans, Phoma hibernica, Rhizopus stolonifer, Trichoderma viride.
Winter SeasonCladosporium cladosprioidesAspergillus niger, Aureobasidium pullulans Gloeosporium sp.Aspergillus flavusColletotrichum falcatum, Penicillium chrysogenum, Rhizopus stoloniferAspergillus sulphureus, A. terreus, Arthrinium cuspidatum, Alternaria alternata, Curvularia pallescens, Epicoccum nigrum, Fusarium semitectum, Mucor hiemalis, Nigrospora sphaerica, Penicillium funiculosum, Phoma hibernica, Sterile mycelium, Trichoderma viride
Summer SeasonNoneFusarium equiseti, Fusarium semitectum, Gloeosporium sp.Aspergillus niger, Fusarium lateritium, Myrothecium roridum, sterile myceliumAspergillus flavus, A. fumigatus, A. sulphureus, Alternaria alternate, Collectotrichum falcatum, Aspergillus nidulans, Phoma hibernica.Aspergillus humicola, A. luchuensis, Aureobasidium pullulans, Cladosporium cladosporioides, Curvularia lunata, Fusarium oxysporum, Mucor hiemalis, Rhizoctonia solani, Rhizopus stolonifer, Trichoderma viride
Partially Decomposed Leaves (PD)Rainy SeasonColletotrichum falcatumAspergillus flavus, A. nigerAspergillus fumigatus, A. luchuensisFusarium oxysporum, Penicillium chrysogenumAlternaria alternata, Choanephora cucurbitarum, Cylindrocarpon sp., Drechslera tetramera, Aspergillus nidulans, Gloeosporium sp., Mucor hiemalis, Penicilium funiculosum, Rhizopus stolonifer, Trichoderma viride.
Winter SeasonCladosporium cladosporioidesAspergillus flavus, Mucor hiemalisAspergillus niger, A. Sulphureus, Aureobasidium pullulans, Gloeosporium sp.,Phoma hibennica, Tnichodenma vinideAlternaria alternate, Collectotrichum falcatum, Curvularia lunata, Fusarium semitectum, Penicillium chrysogenum, Rhizopus stolonifer, sterile mycelium.
Summer SeasonAspergillus fumigatus, A. luchuensis, A. niger Fusarium lateritium, Myrothecium roridum, Absidia repensAlternaria alternateAspergillus flavus, Cladosporium cladosporioides, Aspergillus nidulansAspergillus sulphureus, Aureobasidium pullulans, Collectotrichum falcatum, Monilia geophila, Mucor hiemalis, Penicillium citrinumAspergillus terreus, Curvularia lunata, Fusarium semitectum, Nigrospora sphaerica Rhizopus stolonifer, sterile mycelium, Trichoderma viride
Completely Decomposed Leaves (CD)Rainy SeasonAspergillus niger, Colletotrichum falcatum, Didymium nigripes, Myrothecium roridumAspergillus flavus, Fusarium oxysporumFusarium semitectum, Penicillium chrysogenumAspergillus fumigatus, A. luchuensis, Mucor hiemalis, Rhizopus stolonifer, Trichoderma viride, sterile mycelium.Choanephora cucurbitarum, Aspergillus nidulans, Phaeoramularia graminicola.
Winter SeasonAspergillus flavus, A. Sulphureus, Cladosporium cladosporioidesAspergillus niger, Mucor hiemalisRhizopus stoloniferAlternria alternate, Aspergillus luchuensis, Fusarium semitectum, Memnoniella echinata, sterile mycelium, Trichoderma viride.Stachybotrys atra
Summer SeasonAspergillus flavus, Cladosporium cladosporioides, Fusarium lateritium, Myrothecium roridum, Phoma hibernica, Colletotrichum falcatumAspergillus niger, A. sulphureus, sterile myceliumAspergillus fumigatus, A. luchuensis, Aspergillus nidulans, Fusarium oxysporum, F. semitectumAbsidia repens, Aspergillus terreus, Mucor hiemalis, Penicillium citrinum, Rhizopus stoloniferAspergillus parasiticus, Alternaria alternata, Bipolaris sp., Trichoderma viride.
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Gulati, S.; Chitralekha, P.; Pandit, M.A.; Katyal, R.; Bhandari, N.; Mehta, P.; Rawat, C.D.; Kaur, S.; Kaur, J. Diversity, Succession and Seasonal Variation of Phylloplane Mycoflora of Leucaena leucocephala in Relation to Its Leaf Litter Decomposition. J. Fungi 2022, 8, 608. https://doi.org/10.3390/jof8060608

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Gulati S, Chitralekha P, Pandit MA, Katyal R, Bhandari N, Mehta P, Rawat CD, Kaur S, Kaur J. Diversity, Succession and Seasonal Variation of Phylloplane Mycoflora of Leucaena leucocephala in Relation to Its Leaf Litter Decomposition. Journal of Fungi. 2022; 8(6):608. https://doi.org/10.3390/jof8060608

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Gulati, Saloni, P. Chitralekha, Manisha Arora Pandit, Roma Katyal, Neeru Bhandari, Poonam Mehta, Charu Dogra Rawat, Surinder Kaur, and Jasleen Kaur. 2022. "Diversity, Succession and Seasonal Variation of Phylloplane Mycoflora of Leucaena leucocephala in Relation to Its Leaf Litter Decomposition" Journal of Fungi 8, no. 6: 608. https://doi.org/10.3390/jof8060608

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