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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[3]	Zhang Xi, Wang Guang-En, Li Shao-Qi, Liu Yi, Li Jun-Lan, Qian Yu-Yuan. Transcriptome sequencing-based analysis on the formation mechanism of fiber micronaire differences between two sister lines derived from Gossypium hirsutum-G. barbadense hybrid [J]. Acta Agronomica Sinica, 2026, 52(5): 1442-1458.
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[5]	LI Yi-Qian, XU Shou-Zhen, LIU Ping, MA Qi, XIE Bin, CHEN Hong. Genome-wide association study of yield components using a 40K SNP array and identification of a stable locus for boll weight in upland cotton (Gossypium hirsutum L.) [J]. Acta Agronomica Sinica, 2025, 51(8): 2128-2138.
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[7]	WANG Ya-Wen, QI Zheng-Yang, YOU Jia-Qi, NIE Xin-Hui, CAO Juan, YANG Xi-Yan, TU Li-Li, ZHANG Xian-Long, WANG Mao-Jun. Preparation of cotton 60K functional locus gene chip and its application to genetic research [J]. Acta Agronomica Sinica, 2025, 51(5): 1178-1188.
[8]	DING Jun-Feng, XU Ying-Fei, ZHANG Xiang, CHEN Yuan, CHEN De-Hua. Effects of the plant growth regulator IBA on the survival and growth of substrate- grown transplanted cotton seedlings [J]. Acta Agronomica Sinica, 2025, 51(12): 3331-3341.
[9]	HALIHASHI Yibati, ZHANG Yan, LI Qing-Jun, XU Xin-Peng, HE Ping. Study on smart fertilizer recommendation methods based on yield response and agronomic efficiency for cotton [J]. Acta Agronomica Sinica, 2025, 51(11): 3052-3064.
[10]	ZHAO Hai-Hong, LI Meng-Yuan, LIU Jin-Jing, WANG Yuan-Yuan, DU Lei, WANG Juan, DONG Cheng-Guang, LI Cheng-Qi. Detection of QTNs and QTN-by-environment interactions for plant height in upland cotton (G. hirsutum L.) using the 3VmrMLM method [J]. Acta Agronomica Sinica, 2025, 51(10): 2619-2631.
[11]	LI Ya-Wei, XU Ying-Ying, ZUO Chun-Yang, LIU Ruo-Nan, LIANG Ya-Jun, KONG Jie, ZHANG Xian-Long, MIN Ling. Construction of a meiotic progression identification system in cotton and analysis of its response to high-temperature stress [J]. Acta Agronomica Sinica, 2025, 51(10): 2570-2580.
[12]	CHEN Jia-Wei, LIN Yan, ZHANG Ming-Xing, ZHOU Shi-Jing, RAO Li-Qun, ZHOU Chi, LI Xin. Effects of Bacillus velezensis YCH92 on the rhizosphere microbial community and yield of cotton [J]. Acta Agronomica Sinica, 2025, 51(10): 2821-2835.
[13]	XIE Zhang-Shu, XIE Xue-Fang, TU Xiao-Ju, LIU Ai-Yu, DONG He-Zhong, ZHOU Zhong-Hua. Research progress in phytohormone regulation of square and boll shedding in cotton [J]. Acta Agronomica Sinica, 2025, 51(1): 1-29.
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[15]	LI Chao, FU Xiao-Qiong. Comprehensive evaluation of regional trial varieties of medium mature hybrid cotton in the Yellow River Basin based on GYT biplot [J]. Acta Agronomica Sinica, 2025, 51(1): 30-43.