Isolation and analyses of genes preferentially expressed during early cotton fiber development by subtractive PCR and cDNA array.
Cotton fibers are differentiated epidermal cells originating from the outer integuments of the ovule. To identify genes involved in cotton fiber elongation, we performed subtractive PCR using cDNA prepared from 10 days post anthesis (d.p.a.) wild-type cotton fiber as tester and cDNA from a fuzzless-lintless (fl) mutant as driver. We recovered 280 independent cDNA fragments including most of the previously published cotton fiber-related genes. cDNA macroarrays showed that 172 genes were significantly up-regulated in elongating cotton fibers as confirmed by in situ hybridization in representative cases. Twenty-nine cDNAs, including a putative vacuolar (H+)-ATPase catalytic subunit, a kinesin-like calmodulin binding protein, several arabinogalactan proteins and key enzymes involved in long chain fatty acid biosynthesis, accumulated to greater than 50-fold in 10 d.p.a. fiber cells when compared to that in 0 d.p.a. ovules. Various upstream pathways, such as auxin signal transduction, the MAPK pathway and profilin- and expansin-induced cell wall loosening, were also activated during the fast fiber elongation period. This report constitutes the first systematic analysis of genes involved in cotton fiber development. Our results suggest that a concerted mechanism involving multiple cellular pathways is responsible for cotton fiber elongation.
Ji
Sheng-Jian
SJ
National Laboratory of Protein Engineering and Plant Genetic Engineering and Peking-Yale Joint Center for Plant Molecular Genetics and Agro-Biotechnology, College of Life Sciences, Peking University, Beijing 100871, China.
Lu
Ying-Chun
YC
Feng
Jian-Xun
JX
Wei
Gang
G
Li
Jun
J
Shi
Yong-Hui
YH
Fu
Qiang
Q
Liu
Di
D
Luo
Jing-Chu
JC
Zhu
Yu-Xian
YX
D002455:Cell Division / Q000235:genetics; D003066:Coenzyme A Ligases / Q000235:genetics; D032561:Cotton Fiber; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D003368:Gossypium / Q000235:genetics* / Q000254:growth & development; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012097:Repressor Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA; D000098859:Long-Chain-Fatty-Acid-CoA Ligase
D018076:DNA, Complementary; D012333:RNA, Messenger; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D003066:Coenzyme A Ligases; C090799:FAA2 protein, S cerevisiae; D000098859:Long-Chain-Fatty-Acid-CoA Ligase