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A cotton annexin protein AnxGb6 regulates fiber elongation through its interaction with actin 1.

Huang Y, Wang J, Zhang L, Zuo K - PLoS ONE (2013)

Bottom Line: Here we cloned six Annexin genes (AnxGb) abundantly expressed in fiber from sea-island cotton (G. barbadense). qRT-PCR results indicated that all six G. barbadense annexin genes were expressed in elongating cotton fibers, while only the expression of AnxGb6 was cotton fiber-specific.Analysis of AnxGb6 expression in three cotton genotypes with different fiber length confirmed that AnxGb6 expression was correlated to cotton fiber length, especially fiber elongation rate.Our results demonstrated that AnxGb6 was important for fiber elongation by potentially providing a domain for F-actin organization.

View Article: PubMed Central - PubMed

Affiliation: Plant Biotechnology Research Center, SJTU-Cornell Institute of Sustainable Agriculture and Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.

ABSTRACT
Annexins are assumed to be involved in regulating cotton fiber elongation, but direct evidence remains to be presented. Here we cloned six Annexin genes (AnxGb) abundantly expressed in fiber from sea-island cotton (G. barbadense). qRT-PCR results indicated that all six G. barbadense annexin genes were expressed in elongating cotton fibers, while only the expression of AnxGb6 was cotton fiber-specific. Yeast two hybridization and BiFC analysis revealed that AnxGb6 homodimer interacted with a cotton fiber specific actin GbAct1. Ectopic-expressed AnxGb6 in Arabidopsis enhanced its root elongation without increasing the root cell number. Ectopic AnxGb6 expression resulted in more F-actin accumulation in the basal part of the root cell elongation zone. Analysis of AnxGb6 expression in three cotton genotypes with different fiber length confirmed that AnxGb6 expression was correlated to cotton fiber length, especially fiber elongation rate. Our results demonstrated that AnxGb6 was important for fiber elongation by potentially providing a domain for F-actin organization.

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Real-time quantitative PCR analysis of the annexin genes in Pima-90 and XU142 fl.Expression analysis of annexin genes in G. barbadense (Pima-90) vegetative tissues (R: roots; S: stems; L: leaves), reproductive tissues (C: carpels; −3: ovules in –3 DPA; 0: ovules in 0 DPA; 3: ovules in +3 DPA) and its allele gene expression in G. hirsutum fuzzless-lintless mutant (XU142 fl) reproductive tissues (M −3; ovules in –3 DPA; M0: ovules in 0 DPA; M3: ovules in +3 DPA). The comparative CT method was adopted and the expression was normalized to the levels of Pima-90 and XU142 fl. Error bars represent standard errors.
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pone-0066160-g003: Real-time quantitative PCR analysis of the annexin genes in Pima-90 and XU142 fl.Expression analysis of annexin genes in G. barbadense (Pima-90) vegetative tissues (R: roots; S: stems; L: leaves), reproductive tissues (C: carpels; −3: ovules in –3 DPA; 0: ovules in 0 DPA; 3: ovules in +3 DPA) and its allele gene expression in G. hirsutum fuzzless-lintless mutant (XU142 fl) reproductive tissues (M −3; ovules in –3 DPA; M0: ovules in 0 DPA; M3: ovules in +3 DPA). The comparative CT method was adopted and the expression was normalized to the levels of Pima-90 and XU142 fl. Error bars represent standard errors.

Mentions: Real-time quantitative PCR (qRT-PCR) was performed to investigate the spatial expression patterns of G. barbadense annexin genes in cotton (Figure 3, Figure S1). The qRT-PCR results showed that the expression of all annexin genes was higher in the reproductive tissues than in the vegetative tissues. AnxGb5 and 6 were mainly expressed in the ovule tissues, while very low signals were detected in vegetative tissues (AnxGb6 expression in root), indicating that the functions of AnxGb5 and 6 genes were related to fiber development. In order to get more accurate data for expression pattern of annexins during fiber initials, we performed qRT-PCR to analyze their alleles in G. hirsutum fuzzless-lintless mutant. AnxGb5 and 6 genes had similar expression patterns during fiber initiation, their expression levels were found to constantly increase in the ovules from -3 DPA to 3 DPA. Interestingly, the expression of all 6 annexin genes was found to be significantly higher in the developing fiber cells of Pima-90 in comparison with that of XU142 fiberless mutant, implying a general role for annexins during fiber development and growth (Figure S1).


A cotton annexin protein AnxGb6 regulates fiber elongation through its interaction with actin 1.

Huang Y, Wang J, Zhang L, Zuo K - PLoS ONE (2013)

Real-time quantitative PCR analysis of the annexin genes in Pima-90 and XU142 fl.Expression analysis of annexin genes in G. barbadense (Pima-90) vegetative tissues (R: roots; S: stems; L: leaves), reproductive tissues (C: carpels; −3: ovules in –3 DPA; 0: ovules in 0 DPA; 3: ovules in +3 DPA) and its allele gene expression in G. hirsutum fuzzless-lintless mutant (XU142 fl) reproductive tissues (M −3; ovules in –3 DPA; M0: ovules in 0 DPA; M3: ovules in +3 DPA). The comparative CT method was adopted and the expression was normalized to the levels of Pima-90 and XU142 fl. Error bars represent standard errors.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3672135&req=5

pone-0066160-g003: Real-time quantitative PCR analysis of the annexin genes in Pima-90 and XU142 fl.Expression analysis of annexin genes in G. barbadense (Pima-90) vegetative tissues (R: roots; S: stems; L: leaves), reproductive tissues (C: carpels; −3: ovules in –3 DPA; 0: ovules in 0 DPA; 3: ovules in +3 DPA) and its allele gene expression in G. hirsutum fuzzless-lintless mutant (XU142 fl) reproductive tissues (M −3; ovules in –3 DPA; M0: ovules in 0 DPA; M3: ovules in +3 DPA). The comparative CT method was adopted and the expression was normalized to the levels of Pima-90 and XU142 fl. Error bars represent standard errors.
Mentions: Real-time quantitative PCR (qRT-PCR) was performed to investigate the spatial expression patterns of G. barbadense annexin genes in cotton (Figure 3, Figure S1). The qRT-PCR results showed that the expression of all annexin genes was higher in the reproductive tissues than in the vegetative tissues. AnxGb5 and 6 were mainly expressed in the ovule tissues, while very low signals were detected in vegetative tissues (AnxGb6 expression in root), indicating that the functions of AnxGb5 and 6 genes were related to fiber development. In order to get more accurate data for expression pattern of annexins during fiber initials, we performed qRT-PCR to analyze their alleles in G. hirsutum fuzzless-lintless mutant. AnxGb5 and 6 genes had similar expression patterns during fiber initiation, their expression levels were found to constantly increase in the ovules from -3 DPA to 3 DPA. Interestingly, the expression of all 6 annexin genes was found to be significantly higher in the developing fiber cells of Pima-90 in comparison with that of XU142 fiberless mutant, implying a general role for annexins during fiber development and growth (Figure S1).

Bottom Line: Here we cloned six Annexin genes (AnxGb) abundantly expressed in fiber from sea-island cotton (G. barbadense). qRT-PCR results indicated that all six G. barbadense annexin genes were expressed in elongating cotton fibers, while only the expression of AnxGb6 was cotton fiber-specific.Analysis of AnxGb6 expression in three cotton genotypes with different fiber length confirmed that AnxGb6 expression was correlated to cotton fiber length, especially fiber elongation rate.Our results demonstrated that AnxGb6 was important for fiber elongation by potentially providing a domain for F-actin organization.

View Article: PubMed Central - PubMed

Affiliation: Plant Biotechnology Research Center, SJTU-Cornell Institute of Sustainable Agriculture and Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.

ABSTRACT
Annexins are assumed to be involved in regulating cotton fiber elongation, but direct evidence remains to be presented. Here we cloned six Annexin genes (AnxGb) abundantly expressed in fiber from sea-island cotton (G. barbadense). qRT-PCR results indicated that all six G. barbadense annexin genes were expressed in elongating cotton fibers, while only the expression of AnxGb6 was cotton fiber-specific. Yeast two hybridization and BiFC analysis revealed that AnxGb6 homodimer interacted with a cotton fiber specific actin GbAct1. Ectopic-expressed AnxGb6 in Arabidopsis enhanced its root elongation without increasing the root cell number. Ectopic AnxGb6 expression resulted in more F-actin accumulation in the basal part of the root cell elongation zone. Analysis of AnxGb6 expression in three cotton genotypes with different fiber length confirmed that AnxGb6 expression was correlated to cotton fiber length, especially fiber elongation rate. Our results demonstrated that AnxGb6 was important for fiber elongation by potentially providing a domain for F-actin organization.

Show MeSH
Related in: MedlinePlus