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Acyl-CoA N-acyltransferase influences fertility by regulating lipid metabolism and jasmonic acid biogenesis in cotton.

Fu W, Shen Y, Hao J, Wu J, Ke L, Wu C, Huang K, Luo B, Xu M, Cheng X, Zhou X, Sun J, Xing C, Sun Y - Sci Rep (2015)

Bottom Line: C312) resulted in indehiscent anthers that were full of pollen, diminished filaments and stamens, and plant sterility.We found GhACNAT was involved in lipid metabolism and jasmonic acid (JA) biosynthesis.In GhACNAT-silenced plants, the expression levels of genes involved in lipid metabolism and jasmonic acid biosynthesis were significantly changed, the amount of JA in leaves and reproductive organs was significantly decreased compared with the amounts in C312.

View Article: PubMed Central - PubMed

Affiliation: 1] College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, 310018, Zhejiang, China [2] College of Life and Environmental Science, Hangzhou Normal University, Hangzhou, 310016, Zhejiang, China.

ABSTRACT
Cotton (Gossypium spp.) is an important economic crop and there is obvious heterosis in cotton, fertility has played an important role in this heterosis. However, the genes that exhibit critical roles in anther development and fertility are not well understood. Here, we report an acyl-CoA N-acyltransferase (EC2.3; GhACNAT) that plays a key role in anther development and fertility. Suppression of GhACNAT by virus-induced gene silencing in transgenic cotton (G. hirsutum L. cv. C312) resulted in indehiscent anthers that were full of pollen, diminished filaments and stamens, and plant sterility. We found GhACNAT was involved in lipid metabolism and jasmonic acid (JA) biosynthesis. The genes differentially expressed in GhACNAT-silenced plants and C312 were mainly involved in catalytic activity and transcription regulator activity in lipid metabolism. In GhACNAT-silenced plants, the expression levels of genes involved in lipid metabolism and jasmonic acid biosynthesis were significantly changed, the amount of JA in leaves and reproductive organs was significantly decreased compared with the amounts in C312. Treatments with exogenous methyl jasmonate rescued anther dehiscence and pollen release in GhACNAT-silenced plants and caused self-fertility. The GhACNAT gene may play an important role in controlling cotton fertility by regulating the pathways of lipid synthesis and JA biogenesis.

No MeSH data available.


Related in: MedlinePlus

The morphological changes and GhACNAT expression analysis in transgenic GhACNAT-silenced plants and C312 plants.(a-d) the stamens and stigmas of C312 (a) and transgenic GhACNAT-silenced plants (b, c, d); (e-i) bolls of C312 (e) and transgenic GhACNAT-silenced plants pollinated with C312 pollen (f, h, i); (g) fibers and seeds of C312 and transgenic GhACNAT-silenced plants from left to right. (j) Relative expression levels of GhACNAT measured by qRT-PCR in leaves of GhACNAT-silenced plants (lines 36-2, 36-5, 36-8, 36-10, 36-11, 36-12, 36-16, 36-21 and 36-23) and in C312 plants. The Ubiquitin7 gene (GhUBQ7) was used as an internal control. The values are the means for three replicates (samples of each line collected on days 20, 35 and 50 post-infection). The asterisks indicate statistically significant differences between the transgenic GhACNAT-silenced and WT C312 plants (Significant *P < 0.05, highly significant **P < 0.01, Student’s t-test).
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f3: The morphological changes and GhACNAT expression analysis in transgenic GhACNAT-silenced plants and C312 plants.(a-d) the stamens and stigmas of C312 (a) and transgenic GhACNAT-silenced plants (b, c, d); (e-i) bolls of C312 (e) and transgenic GhACNAT-silenced plants pollinated with C312 pollen (f, h, i); (g) fibers and seeds of C312 and transgenic GhACNAT-silenced plants from left to right. (j) Relative expression levels of GhACNAT measured by qRT-PCR in leaves of GhACNAT-silenced plants (lines 36-2, 36-5, 36-8, 36-10, 36-11, 36-12, 36-16, 36-21 and 36-23) and in C312 plants. The Ubiquitin7 gene (GhUBQ7) was used as an internal control. The values are the means for three replicates (samples of each line collected on days 20, 35 and 50 post-infection). The asterisks indicate statistically significant differences between the transgenic GhACNAT-silenced and WT C312 plants (Significant *P < 0.05, highly significant **P < 0.01, Student’s t-test).

Mentions: There were no obvious differences in the phenotypes of the control plants, the transgenic pCLCrV-PDS plants and the transgenic GhACNAT-silenced plants during vegetative growth except for the mosaic photobleaching spots in the transgenic pCLCrV-PDS plants. In the reproductive stage of the transgenic GhACNAT-silenced plants, the distinct phenotypic changes appeared in the reproductive organs (Fig. 3). The stigmas were forked and warped compared with the stigmas of C312 (Fig. 3a) and the transgenic control plants. The stamens of the transgenic GhACNAT-silenced plants appeared indehiscent but full of viable pollen (Fig. 3b,c,d and 4a,d,g). The filaments became shorter, and the number of stamens decreased significantly (Table S1). No pollen was released from the closed anthers in the transgenic GhACNAT-silenced plants. In the male sterile ms1 mutant, the anthers were closed and shrunken, and no pollen formed (Fig. 4a,f,h). The anthers of the C312 plants were dehiscent and full of active pollen grains, whereas the stamens of the transgenic GhACNAT-silenced plants were indehiscent but as full of viable pollen as the wild-type C312 (Fig. 4d,g). However, the pollen could not be released from the closed stamens. In the transgenic GhACNAT-silenced plants, mature bolls were obtained successfully by pollination with normal C312 pollen. The bolls had two to four loculi containing seeds covered with fibers (Fig. 3e–i).


Acyl-CoA N-acyltransferase influences fertility by regulating lipid metabolism and jasmonic acid biogenesis in cotton.

Fu W, Shen Y, Hao J, Wu J, Ke L, Wu C, Huang K, Luo B, Xu M, Cheng X, Zhou X, Sun J, Xing C, Sun Y - Sci Rep (2015)

The morphological changes and GhACNAT expression analysis in transgenic GhACNAT-silenced plants and C312 plants.(a-d) the stamens and stigmas of C312 (a) and transgenic GhACNAT-silenced plants (b, c, d); (e-i) bolls of C312 (e) and transgenic GhACNAT-silenced plants pollinated with C312 pollen (f, h, i); (g) fibers and seeds of C312 and transgenic GhACNAT-silenced plants from left to right. (j) Relative expression levels of GhACNAT measured by qRT-PCR in leaves of GhACNAT-silenced plants (lines 36-2, 36-5, 36-8, 36-10, 36-11, 36-12, 36-16, 36-21 and 36-23) and in C312 plants. The Ubiquitin7 gene (GhUBQ7) was used as an internal control. The values are the means for three replicates (samples of each line collected on days 20, 35 and 50 post-infection). The asterisks indicate statistically significant differences between the transgenic GhACNAT-silenced and WT C312 plants (Significant *P < 0.05, highly significant **P < 0.01, Student’s t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488762&req=5

f3: The morphological changes and GhACNAT expression analysis in transgenic GhACNAT-silenced plants and C312 plants.(a-d) the stamens and stigmas of C312 (a) and transgenic GhACNAT-silenced plants (b, c, d); (e-i) bolls of C312 (e) and transgenic GhACNAT-silenced plants pollinated with C312 pollen (f, h, i); (g) fibers and seeds of C312 and transgenic GhACNAT-silenced plants from left to right. (j) Relative expression levels of GhACNAT measured by qRT-PCR in leaves of GhACNAT-silenced plants (lines 36-2, 36-5, 36-8, 36-10, 36-11, 36-12, 36-16, 36-21 and 36-23) and in C312 plants. The Ubiquitin7 gene (GhUBQ7) was used as an internal control. The values are the means for three replicates (samples of each line collected on days 20, 35 and 50 post-infection). The asterisks indicate statistically significant differences between the transgenic GhACNAT-silenced and WT C312 plants (Significant *P < 0.05, highly significant **P < 0.01, Student’s t-test).
Mentions: There were no obvious differences in the phenotypes of the control plants, the transgenic pCLCrV-PDS plants and the transgenic GhACNAT-silenced plants during vegetative growth except for the mosaic photobleaching spots in the transgenic pCLCrV-PDS plants. In the reproductive stage of the transgenic GhACNAT-silenced plants, the distinct phenotypic changes appeared in the reproductive organs (Fig. 3). The stigmas were forked and warped compared with the stigmas of C312 (Fig. 3a) and the transgenic control plants. The stamens of the transgenic GhACNAT-silenced plants appeared indehiscent but full of viable pollen (Fig. 3b,c,d and 4a,d,g). The filaments became shorter, and the number of stamens decreased significantly (Table S1). No pollen was released from the closed anthers in the transgenic GhACNAT-silenced plants. In the male sterile ms1 mutant, the anthers were closed and shrunken, and no pollen formed (Fig. 4a,f,h). The anthers of the C312 plants were dehiscent and full of active pollen grains, whereas the stamens of the transgenic GhACNAT-silenced plants were indehiscent but as full of viable pollen as the wild-type C312 (Fig. 4d,g). However, the pollen could not be released from the closed stamens. In the transgenic GhACNAT-silenced plants, mature bolls were obtained successfully by pollination with normal C312 pollen. The bolls had two to four loculi containing seeds covered with fibers (Fig. 3e–i).

Bottom Line: C312) resulted in indehiscent anthers that were full of pollen, diminished filaments and stamens, and plant sterility.We found GhACNAT was involved in lipid metabolism and jasmonic acid (JA) biosynthesis.In GhACNAT-silenced plants, the expression levels of genes involved in lipid metabolism and jasmonic acid biosynthesis were significantly changed, the amount of JA in leaves and reproductive organs was significantly decreased compared with the amounts in C312.

View Article: PubMed Central - PubMed

Affiliation: 1] College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, 310018, Zhejiang, China [2] College of Life and Environmental Science, Hangzhou Normal University, Hangzhou, 310016, Zhejiang, China.

ABSTRACT
Cotton (Gossypium spp.) is an important economic crop and there is obvious heterosis in cotton, fertility has played an important role in this heterosis. However, the genes that exhibit critical roles in anther development and fertility are not well understood. Here, we report an acyl-CoA N-acyltransferase (EC2.3; GhACNAT) that plays a key role in anther development and fertility. Suppression of GhACNAT by virus-induced gene silencing in transgenic cotton (G. hirsutum L. cv. C312) resulted in indehiscent anthers that were full of pollen, diminished filaments and stamens, and plant sterility. We found GhACNAT was involved in lipid metabolism and jasmonic acid (JA) biosynthesis. The genes differentially expressed in GhACNAT-silenced plants and C312 were mainly involved in catalytic activity and transcription regulator activity in lipid metabolism. In GhACNAT-silenced plants, the expression levels of genes involved in lipid metabolism and jasmonic acid biosynthesis were significantly changed, the amount of JA in leaves and reproductive organs was significantly decreased compared with the amounts in C312. Treatments with exogenous methyl jasmonate rescued anther dehiscence and pollen release in GhACNAT-silenced plants and caused self-fertility. The GhACNAT gene may play an important role in controlling cotton fertility by regulating the pathways of lipid synthesis and JA biogenesis.

No MeSH data available.


Related in: MedlinePlus