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Cloning and Functional Analysis of Cotton BI Gene

Author: ZhengLing
Tutor: ZhangJun
School: Shandong Normal University
Course: Developmental Biology
Keywords: cotton BI-1 gene sequence analysis apoptosis expression vector
CLC: S562
Type: Master's thesis
Year: 2011
Downloads: 14
Quote: 0
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Abstract


Bax inhibitor-1 (BI-1) cloned in 1998 was identified a new suppressor genes of apoptosis. Its protein can inhibit apoptosis induced by Bax. In higher plants and animals, BI-1 is an inhibitor of apoptosis, plant which overexpress of BI-1 can increased the tolerability to cell death. BI gene can enhance plant tolerance to drought, salt stress and pathogen infection, which has been proved. That BI gene can enhance cotton resistance to Verticillium wilt is been speculated. This is very meaningful for cultivating new cotton varieties.In this study, using the known part of BI gene sequences of the upland cotton (Gossypium hirsutum L), 4 full-length cDNA sequence was obtained by RACE, which named GhBI-1a1、GhBI-1a2 and GhBI-1b1、GhBI-1b2. Each subgenome of the allotetraploid upland cotton geneme has both GhBI-1a and GhBI-1b and all of which were proved to be expressed in roots and leaves.High homology was showed between amino-acid residues of BI-1a and BI-1b protein, in particular, the carboxyl tail. BI-1a proteins have 5-7 transmembrane regions. Their secondary structures are also similar to other species’s BI-1 protein whose transmembrane regions areα-helix.The expression profiles of the two genes under a variety of stress conditions detected by Q-PCR showed that GhBI-1a and GhBI-1b were induced immediately after stresses and then the expressions were down. It suggests that the two genes are active in the signaling pathways of inhibition apoptosis. It is also suggest that different GhBI-1 genes play a different role under different stresses. GhBI-1a is related with ER stress, but GhBI-1b is related with Verticillium wilt disease.In order to analyze the evolutionary relationships, BI-1 genes are cloned from Gossypium barbadense and the diploid ancestors of allotetroploid cotton, Gossypium arboretum (A subgenome) and Gossypium RaimondiI (D subgenome). From Gossypium barbadense, 4 cDNA sequences BI-1 genes were got, which have high similar with BI-1 from upland cotton. From Gossypium arboretum and Gossypium raimondii, two genes BI-1a and BI-1b were got.In recent years, transgenic arabidopsis and tobacco had proved that BI genes had relation with apoptosis and some biotic non-biotic stresses. To comprehensive understand the biological functions of GhBI-1a and GhBI-1b and the molecular mechanisms of apotosis regulation and stress tolerance, plant expression vectors have been constructed using GhBI-1a and GhBI-1b sequences and transformed into Arabidopsis. RNAi vectors were also been constructed in order to transform cotton to knock-out the corresponding genes.

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CLC: > Agricultural Sciences > Crop > Economic crops > Fiber crops > Cotton
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