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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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Fiber length and AnxGb6 gene expression pattern in Pima-90, Coker312 and T586 cotton seeds.A: Fiber length of Pima-90, Coker312 and T586 cotton seeds at +3, +6, +9 and +12 DPA. Ovules were sectioned and the length of 400 fiber cells was measured under a microscope for each type. Data was processed with Microsoft Excel. Error bars represent standard errors. B: Real-time quantitative PCR analysis of the AnxGb6 gene and its alleles in Pima-90, Coker312 and T586. Expression analysis of AnxGb6 gene and its alleles in Pima-90 (G. barbadense L.), Coker312 (G. hirsutum L.) and T586 (G. hirsutum L.). 3DPA: ovules in +3 DPA, 6DPA: ovules in +6 DPA, 9DPA: ovules in +9 DPA, 12DPA: ovules in +12DPA. The comparative CT method was adopted and the expression was normalized to the levels of Pima-90, Coker312 and T586. Error bars represent standard errors.
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pone-0066160-g008: Fiber length and AnxGb6 gene expression pattern in Pima-90, Coker312 and T586 cotton seeds.A: Fiber length of Pima-90, Coker312 and T586 cotton seeds at +3, +6, +9 and +12 DPA. Ovules were sectioned and the length of 400 fiber cells was measured under a microscope for each type. Data was processed with Microsoft Excel. Error bars represent standard errors. B: Real-time quantitative PCR analysis of the AnxGb6 gene and its alleles in Pima-90, Coker312 and T586. Expression analysis of AnxGb6 gene and its alleles in Pima-90 (G. barbadense L.), Coker312 (G. hirsutum L.) and T586 (G. hirsutum L.). 3DPA: ovules in +3 DPA, 6DPA: ovules in +6 DPA, 9DPA: ovules in +9 DPA, 12DPA: ovules in +12DPA. The comparative CT method was adopted and the expression was normalized to the levels of Pima-90, Coker312 and T586. Error bars represent standard errors.

Mentions: To characterize the effect of AnxGb6 expression on fiber elongation, we chose three cotton varieties with different fiber length including Pima-90 (G. barbadense), Coker312 (G. hirsutum) and T586 (G. hirsutum) for fiber elongation activity analysis. The average fiber lengths of Pima-90, Coker312 and T586 were 33.04±0.28mm, 28.76±0.17mm and 21.09±0.41mm respectively in Shanghai during 2010–2012. At 1 DPA, the fiber cells differentiated and rapidly emerged from the seed-coat surface in all the three cotton cultivars. Thereafter, there was a stage of quick elongation in fiber length (Figure 8A, Table S3). The growth rate of all three cotton varieties was found increased rapidly between 9 and 12 DPA. At 6 DPA, the fiber cells in Pima-90 plants were about 1210 µm long. Six days later (12 DPA), they reached ∼13432 µm in Pima-90 (Figure 8A). The comparison of elongation rates in the 3 tested varieties demonstrated that Pima-90 had the fastest elongation rate followed by Coker 312 and T586. Fiber length of T586 at 6 DPA was only about 200–300 µm, which was equal to fiber length in Pima-90 at 3 DPA. Fiber elongation rate in T586 was 1.5 and 3 fold slower than that in Coker312 and Pima-90 respectively. Previous studies indicate that the elongation time is not significantly different for most of varieties, which is about 20 days for different cotton germplasms with different fiber lengths [69]. Therefore, our results suggested that fiber length was mostly determined by elongation rate and not elongation time.


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)

Fiber length and AnxGb6 gene expression pattern in Pima-90, Coker312 and T586 cotton seeds.A: Fiber length of Pima-90, Coker312 and T586 cotton seeds at +3, +6, +9 and +12 DPA. Ovules were sectioned and the length of 400 fiber cells was measured under a microscope for each type. Data was processed with Microsoft Excel. Error bars represent standard errors. B: Real-time quantitative PCR analysis of the AnxGb6 gene and its alleles in Pima-90, Coker312 and T586. Expression analysis of AnxGb6 gene and its alleles in Pima-90 (G. barbadense L.), Coker312 (G. hirsutum L.) and T586 (G. hirsutum L.). 3DPA: ovules in +3 DPA, 6DPA: ovules in +6 DPA, 9DPA: ovules in +9 DPA, 12DPA: ovules in +12DPA. The comparative CT method was adopted and the expression was normalized to the levels of Pima-90, Coker312 and T586. Error bars represent standard errors.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3672135&req=5

pone-0066160-g008: Fiber length and AnxGb6 gene expression pattern in Pima-90, Coker312 and T586 cotton seeds.A: Fiber length of Pima-90, Coker312 and T586 cotton seeds at +3, +6, +9 and +12 DPA. Ovules were sectioned and the length of 400 fiber cells was measured under a microscope for each type. Data was processed with Microsoft Excel. Error bars represent standard errors. B: Real-time quantitative PCR analysis of the AnxGb6 gene and its alleles in Pima-90, Coker312 and T586. Expression analysis of AnxGb6 gene and its alleles in Pima-90 (G. barbadense L.), Coker312 (G. hirsutum L.) and T586 (G. hirsutum L.). 3DPA: ovules in +3 DPA, 6DPA: ovules in +6 DPA, 9DPA: ovules in +9 DPA, 12DPA: ovules in +12DPA. The comparative CT method was adopted and the expression was normalized to the levels of Pima-90, Coker312 and T586. Error bars represent standard errors.
Mentions: To characterize the effect of AnxGb6 expression on fiber elongation, we chose three cotton varieties with different fiber length including Pima-90 (G. barbadense), Coker312 (G. hirsutum) and T586 (G. hirsutum) for fiber elongation activity analysis. The average fiber lengths of Pima-90, Coker312 and T586 were 33.04±0.28mm, 28.76±0.17mm and 21.09±0.41mm respectively in Shanghai during 2010–2012. At 1 DPA, the fiber cells differentiated and rapidly emerged from the seed-coat surface in all the three cotton cultivars. Thereafter, there was a stage of quick elongation in fiber length (Figure 8A, Table S3). The growth rate of all three cotton varieties was found increased rapidly between 9 and 12 DPA. At 6 DPA, the fiber cells in Pima-90 plants were about 1210 µm long. Six days later (12 DPA), they reached ∼13432 µm in Pima-90 (Figure 8A). The comparison of elongation rates in the 3 tested varieties demonstrated that Pima-90 had the fastest elongation rate followed by Coker 312 and T586. Fiber length of T586 at 6 DPA was only about 200–300 µm, which was equal to fiber length in Pima-90 at 3 DPA. Fiber elongation rate in T586 was 1.5 and 3 fold slower than that in Coker312 and Pima-90 respectively. Previous studies indicate that the elongation time is not significantly different for most of varieties, which is about 20 days for different cotton germplasms with different fiber lengths [69]. Therefore, our results suggested that fiber length was mostly determined by elongation rate and not elongation time.

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