脱落酸对棉花体细胞胚胎发生的影响

doi:10.11963/1002-7807.lpplfg.20201113

摘要/Abstract

摘要：

【目的】研究脱落酸（Abscisic acid, ABA）对棉花体细胞胚胎发生过程中下胚轴脱分化和再分化的影响,优化体细胞胚胎发生体系和初步解析脱落酸调控棉花体细胞胚胎发生分子机制。【方法】以棉花品种中棉所24（CCRI 24）下胚轴为外植体,设置5个ABA浓度0、0.02、0.04、0.06、0.08 μmol·L^-1,分别以A0、A1、A2、A3、A4表示,添加至MSB（MS培养基+B5维生素）培养基诱导愈伤和胚性愈伤,研究ABA对棉花下胚轴初始细胞脱分化、愈伤组织诱导和胚性愈伤组织诱导的影响。【结果】 ABA促进下胚轴初始细胞脱分化;显著提高愈伤组织的脱分化率和增殖率;0.02 μmol·L^-1 ABA显著提高胚性愈伤分化率,0.04~0.08 μmol·L^-1 ABA显著降低胚性愈伤分化率。ABA处理后胚性愈伤和非胚性愈伤的增殖率均显著提高且质地受到影响。0.02~0.08 μmol ABA处理下,LBD和LBD在愈伤起始期上调表达。0.02 μmol·L^-1 ABA处理下,在愈伤增殖早期和中期BBM、LEC1和AGL15上调表达,愈伤增殖后期FUS3、LEA、ABI3基因上调表达。【结论】脱落酸调控的棉花体细胞胚胎发生与相关标记基因的时空性表达密切相关,这些基因表达水平的增加是ABA调控愈伤和胚性愈伤分化的分子基础。

关键词: 陆地棉; 体细胞胚胎发生; 脱落酸; 愈伤组织; 胚性愈伤组织

Abstract:

[Objective] In this study, we analyzed the effect of exogenous hormone abscisic acid (ABA) on cotton callus dedifferentiation and embryogenic callus differentiation during somatic embryogenesis, and aim to optimize the somatic embryogenesis system and to reveal molecular mechanism of ABA regulating somatic embryogenesis. [Method] The hypocotyl of CCRI 24 cotton was used as the explants for callus induction. Callus and embryonic callus were induced by adding different concentrations (0, 0.02, 0.04, 0.06, 0.08 μmol·L^-1) of ABA to MSB medium (MS medium plus vitamin B5) to study the effect of ABA on the initial cellular dedifferentiation, callus induction and embryonal callus induction of cotton hypocotyl. [Result] The initial cellular dedifferentiation was promoted by of 0.02-0.08 μmol·L ^-1 ABA. During the callus induction phase, ABA (0.02-0.08 μmol·L ^-1) significantly increased callus dedifferentiation rate (CDR) and callus proliferation rate (CPR). In the embryogenic callus induction phase, 0.02 μmol·L ^-1 ABA significantly increased embryogenic callus differentiation rate (ECDR), but 0.04-0.08 μmol·L ^-1 ABA treatment significantly inhibited ECDR. ABA treatment (0.02-0.08 μmol·L ^-1) significantly increased embryogenic callus proliferation rate (ECPR) and non-embryogenic callus proliferation rate (N-ECPR), and affected the texture of embryogenic callus and non-embryonic callus. ABA treatment (0.02-0.08 μmol·L ^-1) significantly activated the transcription of LBD16 and LBD29 at the beginning of callus induction. 0.02 μmol·L ^-1 ABA treatment significantly up-regulated the expression levels of BBM, LEC1 and AGL15 at the early and middle stages of callus proliferation, and up-regulated FUS3, LEA, ABI3 expression at the late stage of callus proliferation. [Conclusion] ABA regulated somatic embryogenesis is closely related to the spatiotemporal expression of marker genes, and the increased expression levels of these marker gene is the molecular basis of ABA-mediated differentiation of callus and embryogenic callus.

Key words: upland cotton (Gossypium hirsutum L.); somatic embryogenesis; abscisic acid; callus; embryogenic callus

刘佩佩,魏喜,陈艳丽,王晔,张桂寅,李付广. 脱落酸对棉花体细胞胚胎发生的影响[J]. 棉花学报, 2021, 33(1): 42-53.

Liu Peipei,Wei Xi,Chen Yanli,Wang Ye,Zhang Guiyin,Li Fuguang. Effect of abscisic acid on somatic embryogenesis in upland cotton (Gossypium hirsutum L.)[J]. Cotton Science, 2021, 33(1): 42-53.

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链接本文: http://journal.cricaas.com.cn/Jweb_mhxb/CN/10.11963/1002-7807.lpplfg.20201113

http://journal.cricaas.com.cn/Jweb_mhxb/CN/Y2021/V33/I1/42

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基因名称 Gene name	上游引物 Forward primer (5'-3')	下游引物 Reverse primer (5'-3')
GhHistone3	CTTCAAGACTGATTTGCGTTTCC	GCGCAAAGGTTGGTGTCTTC
GhLBD16	TTGCGCCTTATTTCTGCTCGGA	AATGGTGACAACCGCCTCACAA
GhLBD29	TTCTGCCATGAACAGGGTGCAA	ATCACTGACAGGGAGGTGAGCA
GhBBM	TGTGTACACATCATCGACTG	GCCATATTGCTTGACCTTTG
GhLEC1	CCCCACACGCCAAAATCTCTGA	GATTGATCCACGCTCACCCTCC
GhAGL15	GAAACTTTACGCAGGCAGG	CGAATGTTATGGCTCATCAGAG
GhFUS3	TAGCACGCACGGATTACAGCTC	CATCTTCGTCCGACGCCTTCTT
GhLEA	AGTTCCTTGGTGCTTCTG	CATCGCATATCTTCGTTCC
GhABI3	CAGCAGCAGCCATCTGCTTTTC	CCTCTTTCGTAGCAGACGAGCC