棉花GbSTK基因调控开花和黄萎病抗性的功能研究

doi:10.3724/SP.J.1006.2021.04090

Abstract

Abstract:

Cotton is an important cash crop, and achieving disease resistance as well as early maturity is an important breeding goal. However, the molecular relationship between disease resistance and early maturity of cotton is still a research gap. In previous study, we identified a GbSTK gene response to V. dahliae that could enhance Verticillium wilt resistance in transgenic Arabidopsis. In this study, we found that GbSTK was more highly expressed in resistance and/or tolerant varieties than in susceptible varieties. Knocking down of GbSTK expression in Nongda 601 showed that the silenced plants displayed serious disease symptoms, with disease index from 27.5 (tolerant) to 63.2 (susceptible), suggesting that GbSTK positively regulates cotton Verticillium wilt resistance. More interesting, we found that GbSTK could promote the development of Arabidopsis. At the flowering time point, the average number of rosette leaves in transgenic lines was 7.5, significantly less than the mock plants, and the transgenic plants bloomed five to seven days earlier than the control. The FT and SOC1 gene, regarded as marker genes in Arabidopsis flowering signaling pathway, maintained higher expression level in transgenic lines than in the mock, indicating that GbSTK could accelerate flowering via regulating FT and SOC1 gene expression. We also obtained 30 candidate proteins that could interacted with GbSTK via yeast double hybrid test, which helped to further revealing the molecular regulatory network of GbSTK. Taken together, we first identified a functional gene that could improve disease resistance and early flowering, which would provide a reference for genetic improvement of cotton disease resistant and early maturity.

Key words: cotton, GbSTK, Verticillium wilt resistance, early maturity

CUI Jing, WANG Zhi-Cheng, ZHANG Xin-Yu, KE Hui-Feng, WU Li-Qiang, WANG Xing-Fen, ZHANG Gui-Yin, MA Zhi-Ying, ZHANG Yan. Function study of cotton GbSTK gene in regulating flowering and Verticillium wilt resistance[J].Acta Agronomica Sinica, 2021, 47(1): 30-41.

Figures/Tables 10

Fig. S1

Table 1

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Table 2

Candidate proteins interacting with GbSTK"

编号 No.	蛋白描述 Protein description	功能报道 Function reported	功能分类 Functional classification
1	Cytochrome c oxidase subunit 6b-1	Regulate ROS production^[22]	ROS production
2	Malate dehydrogenase	Related to production of ROS and PCD^[23]	ROS production
3	GTPase-activating protein GYP7	Defense response^[24]	Involved in defense and abiotic/biotic stresses
4	Cytochrome P450 71D8	Defense response^[25]
5	Short-chain dehydrogenase reductase 3b	Involved in plant defense responses^[26]
6	Cysteine proteinase RD21a	Involved in plant defense^[27]
7	Vacuolar protein sorting-associated protein 55	Involved in plant defense^[28]
8	Pathogenesis-related protein STH-2	Defense response^[29]
9	(-)-alpha-terpineol synthase	Involved in defense and abiotic stresses^[30]
10	Glyceraldehyde-3-phosphate dehydrogenase 2	Negatively regulate autophagy and immunity^[31]
11	Phosphoglycerate Ubiquitin-conjugating enzyme E2	Involved in plant immunity^[32]
12	Dirigent protein 16	Responsive to biotic stress^[33]
13	Major allergen Pruar 1	Novel Pathogenesis-related protein^[34]
14	S-Adenosylmethionine synthase	Responsive to abiotic stress^[35]
15	S-adenosylmethionine synthase 2	Responsive to abiotic stress^[36]
16	Probable inorganic phosphate transporter 1-5	Involved in phosphate homeostasis^[37]
17	Glycerophosphodiester phosphodiesterase GDPD1	Response to Pi deficiency^[38]
18	Delta-cadinene synthase isozyme C2	Gossypol biosynthetic^[39]	Secondary metabolism and synthesis
19	UDP-galactose/UDP-glucose transporter 3	Synthesis of cell walls^[40]	Secondary metabolism and synthesis
20	Aldehyde dehydrogenase family 2 member C4	Regulator of the growth-defense trade-off^[41]	Plant development and stress response
21	Probable 2-oxoglutarate-dependent dioxygenase	Related to vascular disease/involved in gibberellin synthesis^[42]
22	Patellin-5	Function in plant development and stress response^[43]
23	Triphosphate tunel metalloenzyme 3	Catalyze diverse enzymatic reactions^[44]
24	Chloroplastic 2,3-bisphosphoglycerate-independent mutase	Involved in photosynthesis and chloroplast development^[45]	Plant growth and development
25	Synaptotagmin-2	Participates in pollen germination and tube growth^[46]	Plant growth and development
26	Mediator of RNA polymerase II transcription subunit	—
27	Alpha-taxilin	—
28	Aconitate hydratase, cytoplasmic	—
29	Putative zinc transporter At3g08650	—
30	40S ribosomal protein S5	—

Table 2

Fig. S2

References 57

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引物名称 Primer name	序列 Primer sequence (5°-3°)	用途 Purpose
STK-F1	CCATATGAAGAATTCAAGCAAATCCTC	基因扩增Gene amplification
STK-R1	GCTGCAGTCAAGGGGCCGTAGGTTGT	基因扩增Gene amplification
VF	ACTAGTGCTGCTCGAACTTCTATTGGGAAG	基因扩增Gene amplification
VR	GGCGCGCCAGGGGCCGTAGGTTGTGTAACTCT	基因扩增Gene amplification
STK-F2 STK-R2	ATGCGGCAACGAACAAGAAT TAGATTGGCACTGAGCTGGT	实时定量qPCR 实时定量qPCR
GhUBQ14-F	CAACGCTCCATCTTGTCCTT	实时定量qPCR
GhUBQ14-R	TGATCGTCTTTCCCGTAAGC	实时定量qPCR
BD-F	TTTGTAATACGACTCACTATAGGGCG	通用载体Vector
BD-R	TTTTCGTTTTAAAACCTAAGAGTCAC	通用载体Vector
T7	TAATACGACTCACTATAGGGCG	通用载体Vector
3°AD	AGATGGTGCACGATGCACAG	通用载体Vector
TUB2-F	ATCCGTGAAGAGTACCCAGAT	实时定量qPCR
TUB2-R	AAGAACCATGCACTCATCAGC	实时定量qPCR
FT-F	CTTGGCAGGCAAACAGTGTATGCAC	实时定量qPCR
FT-R	GCCACTCTCCCTCTGACAATTGTAGA	实时定量qPCR
SOC1-F	AGCTGCAGAAAACGAGAAGCTCTCTG	实时定量qPCR
SOC1-R	GGGCTACTCTCTTCATCACCTCTTCC	实时定量qPCR

Function study of cotton GbSTK gene in regulating flowering and Verticillium wilt resistance

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[4]	ZHENG Shu-Feng, LIU Xiao-Ling, WANG Wei, XU Dao-Qing, KAN Hua-Chun, CHEN Min, LI Shu-Ying. On the green and light-simplified and mechanized cultivation of cotton in a cotton-based double cropping system [J]. Acta Agronomica Sinica, 2022, 48(3): 541-552.
[5]	ZHANG Yan-Bo, WANG Yuan, FENG Gan-Yu, DUAN Hui-Rong, LIU Hai-Ying. QTLs analysis of oil and three main fatty acid contents in cottonseeds [J]. Acta Agronomica Sinica, 2022, 48(2): 380-395.
[6]	ZHANG Te, WANG Mi-Feng, ZHAO Qiang. Effects of DPC and nitrogen fertilizer through drip irrigation on growth and yield in cotton [J]. Acta Agronomica Sinica, 2022, 48(2): 396-409.
[7]	ER Chen, LIN Tao, XIA Wen, ZHANG Hao, XU Gao-Yu, TANG Qiu-Xiang. Coupling effects of irrigation and nitrogen levels on yield, water distribution and nitrate nitrogen residue of machine-harvested cotton [J]. Acta Agronomica Sinica, 2022, 48(2): 497-510.
[8]	ZHAO Wen-Qing, XU Wen-Zheng, YANG Liu-Yan, LIU Yu, ZHOU Zhi-Guo, WANG You-Hua. Different response of cotton leaves to heat stress is closely related to the night starch degradation [J]. Acta Agronomica Sinica, 2021, 47(9): 1680-1689.
[9]	YUE Dan-Dan, HAN Bei, Abid Ullah, ZHANG Xian-Long, YANG Xi-Yan. Fungi diversity analysis of rhizosphere under drought conditions in cotton [J]. Acta Agronomica Sinica, 2021, 47(9): 1806-1815.
[10]	ZENG Zi-Jun, ZENG Yu, YAN Lei, CHENG Jin, JIANG Cun-Cang. Effects of boron deficiency/toxicity on the growth and proline metabolism of cotton seedlings [J]. Acta Agronomica Sinica, 2021, 47(8): 1616-1623.
[11]	GAO Lu, XU Wen-Liang. GhP4H2 encoding a prolyl-4-hydroxylase is involved in regulating cotton fiber development [J]. Acta Agronomica Sinica, 2021, 47(7): 1239-1247.
[12]	MA Huan-Huan, FANG Qi-Di, DING Yuan-Hao, CHI Hua-Bin, ZHANG Xian-Long, MIN Ling. GhMADS7 positively regulates petal development in cotton [J]. Acta Agronomica Sinica, 2021, 47(5): 814-826.
[13]	XU Nai-Yin, ZHAO Su-Qin, ZHANG Fang, FU Xiao-Qiong, YANG Xiao-Ni, QIAO Yin-Tao, SUN Shi-Xian. Retrospective evaluation of cotton varieties nationally registered for the Northwest Inland cotton growing regions based on GYT biplot analysis [J]. Acta Agronomica Sinica, 2021, 47(4): 660-671.
[14]	ZHOU Guan-Tong, LEI Jian-Feng, DAI Pei-Hong, LIU Chao, LI Yue, LIU Xiao-Dong. Efficient screening system of effective sgRNA for cotton CRISPR/Cas9 gene editing [J]. Acta Agronomica Sinica, 2021, 47(3): 427-437.
[15]	HAN Bei, WANG Xu-Wen, LI Bao-Qi, YU Yu, TIAN Qin, YANG Xi-Yan. Association analysis of drought tolerance traits of upland cotton accessions (Gossypium hirsutum L.) [J]. Acta Agronomica Sinica, 2021, 47(3): 438-450.