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Advances in Genetic Engineering and Genome Editing for Crop Improvement

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Plant Sciences".

Deadline for manuscript submissions: closed (30 December 2023) | Viewed by 6780

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Dr. Manoj Kumar

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Institute of Plant Sciences, Agricultural Research Organization, Volcani Center, Rishon LeZion 7505101, Israel
Interests: plant molecular biology; genetic engineering; genome editing; abiotic stress tolerance in plants
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Special Issue Information

Dear Colleagues,

Feeding the ever-increasing global population is a major challenge principally due to expeditious climate changes. Various environmental stressors (abiotic and biotic) may influence growth and development of plant at biochemical, physiological, and molecular levels including changes in gene expression, accumulation of organic solutes, imbalance in phytohormones excretion, and inhibition of plant growth. Conventional plant breeding approach has been carried out for the long time to enhance crop productivity. However, due to its labor intensive and time consuming nature, there is urgent need to adopt new novel biotechnological tools such as genetic engineering and genome editing to develop highly yielding, stress tolerant crop varieties. These novel biotechnological approaches may revolutionize plant breeding and could help secure the global food supply. Genetic engineering is a complex biotechnological tool that manipulates genes at molecular level to develop new varieties with increase yield, resistant to biotic and abiotic stresses, and with enhanced nutritional quality. Genome editing technologies such as transcription-activator-like effector nucleases, zinc finger nucleases, and CRISPR/Cas9 systems have been developed to introduce precise and predictable genome modifications to obtain new crop varieties with desired traits. In this Special Issue of the International Journal of Molecular Sciences, we aim to publish high-quality research articles and reviews on the novel findings on genetic engineering and genome editing approaches to enhance plant growth and stress tolerance for the crop improvement.

Dr. Manoj Kumar
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

genetic engineering
gene expression
plant transformation
genomics
gene editing
CRISPR-Cas
stress responses
agricultural biotechnology
crop protection
crop production

Published Papers (5 papers)

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Research

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11 pages, 18959 KiB

Open AccessArticle

Downregulation of the INDEHISCENT Gene by RNAi Resulted in Desired Pod Shatter Reduction of Lepidium campestre in Subsequent Generations

by Emelie Ivarson, Annelie Ahlman, Jan-Eric Englund, Ida Lager and Li-Hua Zhu

Int. J. Mol. Sci. 2023, 24(21), 15943; https://doi.org/10.3390/ijms242115943 - 3 Nov 2023

Viewed by 555

Abstract

Wild species field cress (Lepidium campestre) has favorable agronomic traits, making it a good candidate for future development as an oil and catch crop. However, the species is very prone to pod shatter, resulting in severe yield losses. This is one of the important agronomic traits that needs to be improved in order to make this species economically viable. In this study, we cloned the L. campestre INDEHISCENT (LcIND) gene and prepared two LcIND-RNAi constructs with the IND promoter (long 400 bp and short 200 bp) from Arabidopsis. A number of stable transgenic lines were developed and evaluated in terms of pod shatter resistance. The majority of the transgenic lines showed increased resistance to pod shatter compared to the wild type, and this resistance was maintained in four subsequent generations. The downregulation of the LcIND gene by RNAi in the transgenic lines was confirmed by qRT-PCR analysis on T₃ lines. Southern blot analysis showed that most of the analyzed lines had a single-copy integration of the transgene, which is desirable for further use. Our results show that it is possible to generate stable transgenic lines with desirable pod shatter resistance by downregulating the LcIND gene using RNAi in field cress, and thus speeding up the domestication process of this wild species. Full article

(This article belongs to the Special Issue Advances in Genetic Engineering and Genome Editing for Crop Improvement)

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22 pages, 5729 KiB

Open AccessArticle

Genome-Wide Analysis of Glycerol-3-Phosphate Acyltransferase (GPAT) Family in Perilla frutescens and Functional Characterization of PfGPAT9 Crucial for Biosynthesis of Storage Oils Rich in High-Value Lipids

by Yali Zhou, Xusheng Huang, Ting Hu, Shuwei Chen, Yao Wang, Xianfei Shi, Miao Yin, Runzhi Li, Jiping Wang and Xiaoyun Jia

Int. J. Mol. Sci. 2023, 24(20), 15106; https://doi.org/10.3390/ijms242015106 - 12 Oct 2023

Cited by 2 | Viewed by 996

Abstract

Glycerol-3-phosphate acyltransferase (GPAT) catalyzes the first step in triacylglycerol (TAG) biosynthesis. However, GPAT members and their functions remain poorly understood in Perilla frutescens, a special edible-medicinal plant with its seed oil rich in polyunsaturated fatty acids (mostly α-linolenic acid, ALA). Here, 14 PfGPATs were identified from the P. frutescens genome and classified into three distinct groups according to their phylogenetic relationships. These 14 PfGPAT genes were distributed unevenly across 11 chromosomes. PfGPAT members within the same subfamily had highly conserved gene structures and four signature functional domains, despite considerable variations detected in these conserved motifs between groups. RNA-seq and RT-qPCR combined with dynamic analysis of oil and FA profiles during seed development indicated that PfGPAT9 may play a crucial role in the biosynthesis and accumulation of seed oil and PUFAs. Ex vivo enzymatic assay using the yeast expression system evidenced that PfGPAT9 had a strong GPAT enzyme activity crucial for TAG assembly and also a high substrate preference for oleic acid (OA, C18:1) and ALA (C18:3). Heterogeneous expression of PfGPAT9 significantly increased total oil and UFA (mostly C18:1 and C18:3) levels in both the seeds and leaves of the transgenic tobacco plants. Moreover, these transgenic tobacco lines exhibited no significant negative effect on other agronomic traits, including plant growth and seed germination rate, as well as other morphological and developmental properties. Collectively, our findings provide important insights into understanding PfGPAT functions, demonstrating that PfGPAT9 is the desirable target in metabolic engineering for increasing storage oil enriched with valuable FA profiles in oilseed crops. Full article

(This article belongs to the Special Issue Advances in Genetic Engineering and Genome Editing for Crop Improvement)

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17 pages, 4521 KiB

Open AccessArticle

CRISPR/Cas9-Mediated HY5 Gene Editing Reduces Growth Inhibition in Chinese Cabbage (Brassica rapa) under ER Stress

by Ye Rin Lee, Ki Seong Ko, Hye Eun Lee, Eun Su Lee, Koeun Han, Jae Yong Yoo, Bich Ngoc Vu, Ha Na Choi, Yoo Na Lee, Jong Chan Hong, Kyun Oh Lee and Do Sun Kim

Int. J. Mol. Sci. 2023, 24(17), 13105; https://doi.org/10.3390/ijms241713105 - 23 Aug 2023

Cited by 1 | Viewed by 1362

Abstract

Various stresses can affect the quality and yield of crops, including vegetables. In this study, CRISPR/Cas9 technology was employed to examine the role of the ELONGATED HYPOCOTYL 5 (HY5) gene in influencing the growth of Chinese cabbage (Brassica rapa). Single guide RNAs (sgRNAs) were designed to target the HY5 gene, and deep-sequencing analysis confirmed the induction of mutations in the bZIP domain of the gene. To investigate the response of Chinese cabbage to endoplasmic reticulum (ER) stress, plants were treated with tunicamycin (TM). Both wild-type and hy5 mutant plants showed increased growth inhibition with increasing TM concentration. However, the hy5 mutant plants displayed less severe growth inhibition compared to the wild type. Using nitroblue tetrazolium (NBT) and 3,3′-diaminobenzidine (DAB) staining methods, we determined the amount of reactive oxygen species (ROS) produced under ER stress conditions, and found that the hy5 mutant plants generated lower levels of ROS compared to the wild type. Under ER stress conditions, the hy5 mutant plants exhibited lower expression levels of UPR- and cell death-related genes than the wild type. These results indicate that CRISPR/Cas9-mediated editing of the HY5 gene can mitigate growth inhibition in Chinese cabbage under stresses, improving the quality and yield of crops. Full article

(This article belongs to the Special Issue Advances in Genetic Engineering and Genome Editing for Crop Improvement)

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14 pages, 1717 KiB

Open AccessArticle

Overexpression of the Purple Perilla (Perilla frutescens (L.)) FAD3a Gene Enhances Salt Tolerance in Soybean

by Zhan Li, Ying Wang, Lili Yu, Yongzhe Gu, Lijuan Zhang, Jun Wang and Lijuan Qiu

Int. J. Mol. Sci. 2023, 24(13), 10533; https://doi.org/10.3390/ijms241310533 - 23 Jun 2023

Cited by 1 | Viewed by 1082

Abstract

The increasingly serious trend of soil salinization inhibits the normal growth and development of soybeans, leading to reduced yields and a serious threat to global crop production. Microsomal ω-3 fatty acid desaturase encoded by the FAD3 gene is a plant enzyme that plays a significant role in α-linolenic acid synthesis via regulating the membrane fluidity to better accommodate various abiotic stresses. In this study, PfFAD3a was isolated from perilla and overexpressed in soybeans driven by CaMV P35S, and the salt tolerance of transgenic plants was then evaluated. The results showed that overexpression of PfFAD3a increased the expression of PfFAD3a in both the leaves and seeds of transgenic soybean plants, and α-linolenic acid content also significantly increased; hence, it was shown to significantly enhance the salt tolerance of transgenic plants. Physiological and biochemical analysis showed that overexpression of PfFAD3a increased the relative chlorophyll content and PSII maximum photochemical efficiency of transgenic soybean plants under salt stress; meanwhile, a decreased accumulation of MDA, H₂O₂, and

O_{2}^{• -}

, increased the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbic acid peroxidase (APX), as well as the production of proline and soluble sugar. In summary, the overexpression of PfFAD3a may enhance the salt tolerance in transgenic soybean plants through enhanced membrane fluidity and through the antioxidant capacity induced by C18:3. Full article

(This article belongs to the Special Issue Advances in Genetic Engineering and Genome Editing for Crop Improvement)

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Review

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18 pages, 992 KiB

Open AccessReview

Transgenic Improvement for Biotic Resistance of Crops

by Haoqiang Yu, Yingge Wang, Fengling Fu and Wanchen Li

Int. J. Mol. Sci. 2022, 23(22), 14370; https://doi.org/10.3390/ijms232214370 - 19 Nov 2022

Cited by 2 | Viewed by 2064

Abstract

Biotic constraints, including pathogenic fungi, viruses and bacteria, herbivory insects, as well as parasitic nematodes, cause significant yield loss and quality deterioration of crops. The effect of conventional management of these biotic constraints is limited. The advances in transgenic technologies provide a direct and directional approach to improve crops for biotic resistance. More than a hundred transgenic events and hundreds of cultivars resistant to herbivory insects, pathogenic viruses, and fungi have been developed by the heterologous expression of exogenous genes and RNAi, authorized for cultivation and market, and resulted in a significant reduction in yield loss and quality deterioration. However, the exploration of transgenic improvement for resistance to bacteria and nematodes by overexpression of endogenous genes and RNAi remains at the testing stage. Recent advances in RNAi and CRISPR/Cas technologies open up possibilities to improve the resistance of crops to pathogenic bacteria and plant parasitic nematodes, as well as other biotic constraints. Full article

(This article belongs to the Special Issue Advances in Genetic Engineering and Genome Editing for Crop Improvement)

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