Isolation of genes preferentially expressed in cotton fibers by cDNA filter arrays and RT-PCR

Abstract

Cotton fibers are single-cell trichomes derived from the outer integument cells of ovules. Recent studies showed that the rapid elongation of fibers was coordinated with the expression of a subset of fiber-specific genes. Here we report the use of filter arrays of cDNAs to identify genes preferentially expressed in cotton fibers. An array of 1536 clones was hybridized with cDNA probes prepared from wild type (fiber-containing) and fl mutant (naked) ovules at 5 days post-anthesis (DPA), respectively. Subsequent RT-PCR analyses of 14 genes identified ten that were highly transcribed in cotton fibers. Among them, an RD22-like protein (GhRDL ), a putative acyltransferase (GhACY ), a Fiddlehead homolog (GhFDH ), a serine carboxypeptidase-like protein (GhSCP ), two tubulin components (GhTUA6 and GhTUB1 ) and the previously reported gene encoding fiber protein E6, showed a fiberenriched expression pattern. The other three genes, including an actin (GhACT ), a putative cellulose synthase catalytic subunit (GhCesA-5 ) and a putative 24-sterol-C -methyltransferase (GhSMT ), were actively transcribed in fibers during the elongation stage, but their transcripts were also clearly present in other tissues. The possible roles of these proteins in cotton fiber development and growth are discussed. #

Control of Plant Trichome Development by a Cotton Fiber MYB Gene[W]

“…Our previous study showed that transcripts of a cotton RDL gene were enriched in developing fiber cells (Li et al, 2002). DNA gel blot hybridization revealed two RDL genes, namely RDL1 and RDL2, in the genome of G. arboreum (see Supplemental Figure 1 online), an A-genome diploid cotton cultivar (Rong et al, 2004).…”

“…Analyses of gene expression patterns have identified numerous genes whose transcripts are enriched in fiber cells (John and Crow, 1992;Li et al, 2002;Ji et al, 2003), including those encoding metabolism enzymes that are essential to cell growth (Pear et al, 1996;Ruan et al, 2003). For transcription factors, a noteworthy work is the isolation of six MYB genes from G. hirsutum (Loguerico et al, 1999;Cedroni et al, 2003).…”

Gene expression and metabolite profiles of cotton fiber during cell elongation and secondary cell wall synthesis

“…The quality of fibers npg isolated from the 3 DPA ovules was also checked under a microscope. Total RNA was isolated from the -3 to 5 DPA ovules and the 6 to 24 DPA fibers by a cold-phenol method as described previously [27,28]. Poly (A)+RNA was purified by an Oligotex mRNA Midi kit (Qiagen, Valencia, CA, USA) according to the manufacturer's instruction, and cDNA libraries were constructed following the ZAPcDNA Synthesis kit (Stratagene, La Jolla, CA, USA).…”

“…Further investigation by KOBAS [27] mapped 1 116 out of the 5 122 cotton ESTs to 164 pathways, and 152 stage-differential genes to 85 pathways. After hyper geometric distribution and FDR corrections, seven metabolic pathways involving secondary metabolite, fatty acid and carbohydrate metabolisms show significant changes (q < 0.05) during cotton fiber development (Table 2).…”

Trichome Specific Expression: Promoters and Their Applications

“…These early studies were followed by genomics approaches, including Expressed Sequence Tag (EST) library sequencing and microarray hybridization. In particular, EST libraries corresponding to various stages of development were produced and these provide invaluable resources for the identification of fiber specific genes (Li et al, 2002a;Arpat et al, 2004;Udall et al, 2006;Yang et al, 2006). As genes from these EST collections start being characterized, more information has become available on the pattern of expression and the importance of some transcription factors in fiber development (Lee et al, 2007).…”