Function Analysis of GhLhcb2A1 Gene in Response to Low Temperature and Drought in Upland Cotton

doi:10.7668/hbnxb.20195341

Abstract

Abstract:

Light-harvesting chlorophyll a/b binding proteins are important in plant photosynthesis and abiotic stress response.To study the characteristics of GhLhcb2A1 and its expression patterns and functions in low temperature and drought response in upland cotton,full-length CDS of GhLhcb2A1 gene was cloned from the leaf cDNA of Jimian 262 by PCR.Bioinformatic analysis was conducted to learn the basic characteristics of the gene.The expression patterns and functions in low temperature and drought response were evaluated by qRT-PCR and virus-induced gene silencing.It was shown that the length of GhLhcb2A1 CDS was 798 bp,encoding 265 amino acids.GhLhcb2A1 was highly expressed in leaves and was significantly up-regulated in leaves and roots under low temperature and drought treatment.Compared with the control,its expression maximized at 3 h under low temperature and drought in leaves with 17.42 and 30.03 folds increase respectively,whereas maximized at 6 h under low temperature and 12 h under drought in roots with 11.65 and 65.04 folds respectively.Subcellular localization assay verified that GhLhcb2A1 was expressed in the chloroplasts of cells.Compared with the control plants,GhLhcb2A1 silenced plants showed a more severe phenotype of water loss and dryness under both low temperatures and drought.The accumulated malondialdehyde content in the leaves of the silenced plants was significantly higher than that of the control,while the proline content and superoxide dismutase activity were significantly lower than those of the controls,suggesting that GhLhcb2A1 silenced plant reduced the resistance to low temperature and drought.The above results implied that this gene played a positive role in regulating low temperature and drought response.

Key words: Upland cotton, GhLhcb2A1, Low temperature, Drought, Light-harvesting chlorophyll a/b binding protein

CAI Xiao, LIU Cunjing, ZHANG Sujun, LI Xinghe, WANG Haitao, TANG Liyuan, ZHANG Jianhong. Function Analysis of GhLhcb2A1 Gene in Response to Low Temperature and Drought in Upland Cotton[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(1): 14-21. doi: 10.7668/hbnxb.20195341.

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Figures/Tables 6

Tab.1 Primer sequences

引物名称 Prime name	序列(5'—3') Sequence (5'—3')	用途 Function
GhLhcb2A1-F	ATGGCAACCTCTGCTATCCA	基因克隆
GhLhcb2A1-R	TCATTTTCCGGGGACAAAGT
GhLhcb2A1-q-F	GTTGGCCAGACCGCTTTGAA	实时荧光定量PCR
GhLhcb2A1-q-R	CATGGTAACACGACCACGATTG
GhLhcb2A1-GFP-F	TTCCCGGGATGGCAACCTCTGCTATCCA	GFP融合载体构建
GhLhcb2A1-GFP-R	TTGGATCCTTTTCCGGGGACAAAGTTAG
GhUBQ7-F	GAAGGCATTCCACCTGACCAAC	实时荧光定量PCR
GhUBQ7-R	CTTGACCTTCTTCTTCTTGTGCTTG

Fig.1 Prediction of phosphorylation sites and transmembrane regions in GhLhcb2A1 protein

Fig.2 Tertiary structure of GhLhcb2A1 protein

Fig.3 Expression of GhLhcb2A1 gene in different tissues,low temperature and drought response The least significant difference(LSD)test was used,lowercase letters represent significant differences between samples(P<0.05).The same as Fig.5.

Fig.4 Subcellular localization of GhLhcb2A1 protein

Fig.5 Response of GhLhcb2A1 silencing plants to low temperature and drought A.Positive control of GhCLA1 silencing plants showed green loss in the leaf;B.Expression analysis of GhLhcb2A1 gene in VIGS plant leaves by qRT-PCR;C.Phenotype of GhLhcb2A1 silencing plants and negative control plants under stress at 0 h;D.Phenotype of GhLhcb2A1 silencing plants and negative control plants after 7 d under cold stress;E.Phenotype of GhLhcb2A1 silencing plants and negative control plants after 10 d under drought stress.

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