甘蔗节间伸长过程赤霉素生物合成关键基因的表达及相关植物激素动态变化

doi:10.11983/CBB18139

植物学报 ›› 2019, Vol. 54 ›› Issue (4): 486-496.DOI: 10.11983/CBB18139

甘蔗节间伸长过程赤霉素生物合成关键基因的表达及相关植物激素动态变化

范业赓¹,丘立杭¹,黄杏¹,周慧文¹,甘崇琨²,李杨瑞¹,杨荣仲¹,吴建明^1,^*(),陈荣发^1,^*()

1 广西壮族自治区农业科学院, 甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室, 南宁 530007
2 广西崇左市江州区土肥站, 崇左 532200

收稿日期:2018-06-19 接受日期:2018-12-10 出版日期:2019-07-10 发布日期:2020-01-08
通讯作者: 吴建明,陈荣发
基金资助:
国家自然科学基金(31360312);国家自然科学基金(31701363);广西自然科学基金(2016GXNSFBA380034);广西自然科学基金(2017GXNSFBA198050);广西自然科学基金(2018GXNSFAA138149);广西科技计划(桂科攻1598006-1-2E);广西科技计划(桂科AA17202042-13);国家农业产业技术体系广西甘蔗创新团队专项(gjnytxgxcxtd-03)

Expression Analysis of Key Genes in Gibberellin Biosynthesis and Related Phytohormonal Dynamics During Sugarcane Internode Elongation

Yegeng Fan¹,Lihang Qiu¹,Xing Huang¹,Huiwen Zhou¹,Chongkun Gan²,Yangrui Li¹,Rongzhong Yang¹,Jianming Wu^1,^*(),Rongfa Chen^1,^*()

1 Key Laboratory of Sugarcane Biotechnology and Genetic Improvement, Ministry of Agriculture/Guangxi Key Laboratory of Sugarcane Genetic Improvement, Sugarcane Research Institute, Guangxi Zhuang Autonomous Region Academy of Agricultural Sciences, Nanning 530007, China
2 Chongzuo Jiangzhou Soil Fertility Station, Chongzuo 532200, China

Received:2018-06-19 Accepted:2018-12-10 Online:2019-07-10 Published:2020-01-08
Contact: Jianming Wu,Rongfa Chen

摘要/Abstract

摘要： 以甘蔗(Saccharum officinarum)优良品种桂糖42号(GT42)为研究材料, 分别于未伸长期(9-10叶龄以前) (Ls1)、伸长初期(12-13叶龄) (Ls2)和伸长盛期(15-16叶龄) (Ls3)取甘蔗第2片真叶(自顶部起)对应的节间组织, 测定其赤霉素(GA)、生长素(IAA)、油菜素甾醇(BR)、细胞分裂素(CTK)、乙烯(ETH)和脱落酸(ABA)的含量, 并通过实时荧光定量PCR (qRT-PCR)分析赤霉素合成途径关键基因GA₂₀氧化酶基因(GA20-Oxidase1)、赤霉素受体基因(GID1)和DELLA蛋白编码基因(GAI)的差异表达。结果表明, 在甘蔗伸长期间, GA和IAA含量呈现上升趋势, CTK和ABA含量呈下降趋势, ETH含量先上升后下降, BR含量则变化不明显; GA20-Oxidase1和GID1的表达呈上升趋势, 而GAI的表达则呈下降趋势, 这与相关植物激素的变化基本一致。综上, 甘蔗节间伸长过程主要与GA和IAA相关, 其次为CTK和ABA, 而ETH受到IAA的调控影响节间伸长; 植物激素间通过相互作用调控GA20-Oxidase1、GID1和GAI的表达, 影响GA含量和GA的信号转导过程, 进而影响甘蔗节间的伸长。该研究揭示了甘蔗节间伸长过程中赤霉素生物合成途径和信号转导关键基因的差异表达及植物激素含量的动态变化规律。

关键词: 植物激素, 赤霉素, 基因表达分析, 节间伸长, 甘蔗

Abstract: In this study, Saccharum officinarum GT42 was used as the research material. Second internode tissue of sugarcane at the non-elongation stage (Ls1), early elongation stage (Ls2) and elongation stage (Ls3) was collected to measure plant hormones such as gibberellic acid (GA), indole acetic acid (IAA), brassinosteroids (BR), cytokinin (CTK), ethylene (ETH) and abscisic acid (ABA). The differential expression of the key genes GA20-Oxidase1, GID1 and GAI was analyzed by qRT-PCR. During the elongation stage, the contents of GA and IAA showed an upward trend, but CTK and ABA contents showed a downward trend. ETH content was increased at first and then decreased, whereas BR content did not change. The expression of GA20-Oxidase 1 and GID1 increased, and that of GAI decreased, which was closely related to the changes in GA content. Therefore, we considered that the internode elongation of sugarcane is mainly related to GA and IAA, then are CTK and ABA, and ETH is regulated by IAA to affect internode elongation. The interaction between these phytohormones regulates the expression of GA20-Oxidase 1, GID1 and GAI, which affects the content and signal transduction of GA, thereby affecting the internode elongation of sugarcane. The study clarifies the differential expression of key genes in the gibberellin biosynthesis pathway and signal transduction process, along with the dynamic changes of plant hormone content during internode elongation of sugarcane.

Key words: phytohormone, gibberellin, gene expression analysis, internode elongation, sugarcane

范业赓,丘立杭,黄杏,周慧文,甘崇琨,李杨瑞,杨荣仲,吴建明,陈荣发. 甘蔗节间伸长过程赤霉素生物合成关键基因的表达及相关植物激素动态变化. 植物学报, 2019, 54(4): 486-496.

Yegeng Fan,Lihang Qiu,Xing Huang,Huiwen Zhou,Chongkun Gan,Yangrui Li,Rongzhong Yang,Jianming Wu,Rongfa Chen. Expression Analysis of Key Genes in Gibberellin Biosynthesis and Related Phytohormonal Dynamics During Sugarcane Internode Elongation. Chinese Bulletin of Botany, 2019, 54(4): 486-496.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB18139

https://www.chinbullbotany.com/CN/Y2019/V54/I4/486

图/表 5

表1 实时荧光定量PCR引物序列

Table 1 Primers for real-time quantitative PCR

Genes	Sequences of primers (5'-3')	Tm (°C)	Product length (bp)
25S rRNA	F: CGAATACGAACCGTGAAAGC	58.4	164
25S rRNA	R: ACAATGATAGGAAGAGCCGACA	59.0	164
GA20-Oxidase1	F: CCGTCGTGGACTACTTCG	54.0	205
GA20-Oxidase1	R: CACGGCGGGTAGTAGTTGAG	58.3	205
GID1	F: GGTCATCCTCTTCTTCCACGG	60.8	119
GID1	R: AGTTGACGGACACCACGACG	61.3	119
GAI	F: GCGACGACAAGGATGAGGA	58.8	186
GAI	R: TGAAGCGGTCAAGGAATGTG	59.2	186

表2 不同时期甘蔗的伸长情况(平均值±标准差)

Table 2 The elongation of sugarcane at different stages (means±SD)

Period	Plant height (cm)	Elongation rate (cm·d^-1)	Internode number	Average internode length (cm)
Ls1	39.38±1.10	0.44±0.01	4±1	6.09±0.28
Ls2	68.37±1.25**	0.97±0.07**	7±1**	8.05±0.18**
Ls3	147.13±0.85**	2.63±0.06**	12±1**	11.43±0.07**

图1 不同时期甘蔗节间生长的差异 (A) 未伸长期(Ls1)甘蔗株高; (B) 伸长初期(Ls2)甘蔗株高; (C) 伸长盛期(Ls3)甘蔗株高。Bars=10 cm

Figure 1 Changes of sugarcane elongation at different stages (A) The sugarcane plant height of non-elongation stage (Ls1); (B) The sugarcane plant height of early elongation stage (Ls2); (C) The sugarcane plant height of elongation stage (Ls3). Bars=10 cm

图2 不同伸长时期甘蔗节间植物激素含量变化 (A) 生长素(IAA); (B) 油菜素甾醇(BR); (C) 细胞分裂素(CTK); (D) 脱落酸(ABA); (E) 乙烯(ETH)。不同小写字母表示差异显著(P<0.05), 不同大写字母表示差异极显著(P<0.01)。Ls1-Ls3同表2。

Figure 2 Changes of endogenous hormone contents in sugarcane internode during different elongation stages (A) Indole acetic acid (IAA); (B) Brassinosteroids (BR); (C) Cytokinin (CTK); (D) Abscisic acid (ABA); (E) Ethylene (ETH). The different lowercase letters indicate significant differences (P<0.05), and the different capital letters indicate extremely significant differences (P<0.01). Ls1-Ls3 see Table 2.

图3 甘蔗节间伸长过程中赤霉素含量变化及其合成途径关键基因的差异表达 (A) 赤霉素(GA)含量; (B) GA20-Oxidase1的表达量; (C) GID1的表达量; (D) GAI的表达量。不同小写字母表示差异显著(P<0.05), 不同大写字母表示差异极显著(P<0.01)。Ls1-Ls3同表2。

Figure 3 The changes of gibberellin content and genes differential expression of synthetic pathways during sugarcane internode elongation (A) The content of gibberellic acid (GA); (B) The expression level of GA20-Oxidase1; (C) The expression level of GID1; (D) The expression level of GAI. The different lowercase letters indicate significant differences (P<0.05), and the different capital letters indicate extremely significant differences (P<0.01). Ls1-Ls3 see Table 2.

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甘蔗节间伸长过程赤霉素生物合成关键基因的表达及相关植物激素动态变化

Expression Analysis of Key Genes in Gibberellin Biosynthesis and Related Phytohormonal Dynamics During Sugarcane Internode Elongation

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