Characterization of JsWOX1 and JsWOX4 during Callus and Root Induction in the Shrub Species Jasminum sambac
Abstract
:1. Introduction
2. Results
2.1. Establishment of J. sambac Callus Culture and the Role of Auxin and Cytokinin
2.2. Analysis of WOX Gene Expression during Callus Induction
2.3. In Situ Detection of JsWOX1 and JsWOX4 Transcripts in Calli
2.4. Effects of Ectopic Expression of JsWOX1 and JsWOX4 on Callus Morphogenesis and Gene Expressions
3. Discussion
4. Materials and Methods
4.1. Plant Materials and Culture Conditions
4.2. Transverse Sections, Histological Observation and Scanning Electron Microscopy (SEM)
4.3. Full-Length cDNA Cloning and Agrobacterium-Mediated Transformation
4.4. In situ mRNA Hybridization
4.5. Quantitative RT-PCR
4.6. Sequence Alignment and Phylogenetic Tree
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Treatment | WPM Medium + NAA + BA (M) | Ratio NAA/BA | Percent Explant with Callus (%) (1 Month) | Percent Explant with Rooted Callus (%) (2 Months) | Extended Callus Culture (up to 6 months) | |
---|---|---|---|---|---|---|
1 | 8.06 | 0 | 100/0 | 80 (n = 22) | 0 | browning, dying |
2 | 6.04 | 1.67 | 78/22 | 83 (n = 20) | 0 | browning, dying |
3 | 4.03 | 3.33 | 55/45 | 85 (n = 30) | 0 | browning, dying |
4 | 2.01 | 5.00 | 29/71 | 65 (n = 18) | 0 | browning, dying |
5 | 0 | 6.66 | 0/100 | 15 (n = 18) | 0 | browning, dying |
6 | 2.69 | 8.88 | 23/77 | 86 (n = 400) | 3 | most browning |
7 | 1.07 | 8.88 | 11/89 | 98 (n = 450) | 6 | pale green, stable |
8 | 0 | 8.88 | 0/100 | 10 (n = 400) | 1 | browning, dying |
9 | 16.11 | 0 | 100/0 | 86 (n = 23) | 13 | some browning |
10 | 12.08 | 3.33 | 78/22 | 96 (n = 24) | 12.5 | browning |
11 | 8.06 | 6.66 | 55/45 | 98 (n = 23) | 13 | dark green, stable |
12 | 4.03 | 9.99 | 29/71 | 98 (n = 126) | 30 | pale green, stable |
13 | 0 | 13.32 | 0/100 | 40 (n = 23) | 0 | browning, dying |
14 | 48.34 | 0 | 100/0 | 60 (n = 19) | 0 | browning, dying |
15 | 36.25 | 9.99 | 78/22 | 68 (n = 18) | 0 | browning, dying |
16 | 24.17 | 19.98 | 55/45 | 85 (n = 25) | 4 | some browning |
17 | 12.08 | 29.97 | 29/71 | 34 (n = 29) | 7 | some browning |
18 | 0 | 39.96 | 0/100 | 10 (n = 23) | 0 | browning, dying |
19 | 112.78 | 0 | 100/0 | 14 (n = 13) | 0 | browning, dying |
20 | 84.59 | 23.31 | 78/22 | 15 (n = 18) | 0 | browning, dying |
21 | 56.39 | 46.63 | 55/45 | 8 (n = 14) | 0 | browning, dying |
22 | 28.20 | 69.94 | 29/71 | 40 (n = 22) | 4.5 | browning, dying |
23 | 0 | 93.25 | 0/100 | 4 (n = 10) | 0 | browning, dying |
Gene | Forward (5′-3′) | Reverse (5′-3′) |
---|---|---|
JsWOX1 | ACACCGTCGGCTGATCAAAT | ATTCCAGTTGACGTCGCCTT |
JsWOX4 | TAGAGGAGGAATGCGAACGC | TTCTGTCTCTCGCGTGCTTT |
JsWOX13x1 | ATTACGGGCAGGCAGAGATG | CCGTGTTGGCATAACTCTGC |
JsWOX13x2 | TGTACCCTGGTGGCCATAGA | TCTGCTTGCTTGGAGTTCCA |
c92402 | TTTTCGCAGCTTGCTTCCAC | CGGGACCCAACGATGAGAAT |
c76574 | GCTCAAGTGGGCATGGGATT | GCACCTCTCTCGTATCGTGT |
c19299 | GCTGTGTCTAGGTGCATGGT | TCCGCTTCATCGTACACTCC |
c16725 | AAAGTTGTGGTGGAGTGGCA | CGACAGTCCCGAAACCAAGA |
JsActin2 | TCTCTATGGTAACATTGTCCTG | TCTCTATGGTAACATTGTCCTG |
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Lu, Y.; Liu, Z.; Lyu, M.; Yuan, Y.; Wu, B. Characterization of JsWOX1 and JsWOX4 during Callus and Root Induction in the Shrub Species Jasminum sambac. Plants 2019, 8, 79. https://doi.org/10.3390/plants8040079
Lu Y, Liu Z, Lyu M, Yuan Y, Wu B. Characterization of JsWOX1 and JsWOX4 during Callus and Root Induction in the Shrub Species Jasminum sambac. Plants. 2019; 8(4):79. https://doi.org/10.3390/plants8040079
Chicago/Turabian StyleLu, Ying, Zhuoyi Liu, Meiling Lyu, Yuan Yuan, and Binghua Wu. 2019. "Characterization of JsWOX1 and JsWOX4 during Callus and Root Induction in the Shrub Species Jasminum sambac" Plants 8, no. 4: 79. https://doi.org/10.3390/plants8040079