Abstract
Analysis of mutants and gene expression patterns provides a powerful approach for investigating genes involved in key stages of plant fiber development. In this study, lintless-fuzzless XinWX and linted-fuzzless XinFLM with a single genetic locus difference for lint were used to identify differentially expressed genes. Scanning electron microscopy showed fiber initiation in XinFLM at 0 days post anthesis (DPA). Fiber transcriptional profiling of the lines at three initiation developmental stages (-1, 0, 1 DPA) was performed using an oligonucleotide microarray. Loop comparisons of the differentially expressed genes within and between the lines was carried out, and functional classification and enrichment analysis showed that gene expression patterns during fiber initiation were heavily associated with hormone metabolism, transcription factor regulation, lipid transport, and asparagine biosynthetic processes, as previously reported. Further, four members of the allene-oxide cyclase (AOC) family that function in jasmonate biosynthesis were parallel up-regulation in fiber initiation, especially at -1 DPA, compared to other tissues and organs in linted-fuzzed TM-1. Real time-quantitative PCR (RT-qPCR) analysis in different fiber mutant lines revealed that AOCs were up-regulated higher at -1 DPA in lintless-fuzzless than that in linted-fuzzless and linted-fuzzed materials, and transcription of the AOCs was increased under jasmonic acid (JA) treatment. Expression analysis of JA biosynthesis-associated genes between XinWX and XinFLM showed that they were up-regulated during fiber initiation in the fuzzless-lintless mutant. Taken together, jasmonic acid-associated metabolism was related to cotton fiber initiation. Parallel up-regulation of AOCs expression may be important for normal fiber initiation development, while overproduction of AOCs might disrupt normal fiber development.
Highlights
Gossypium hirsutum is an important economic crop and is a major source of both natural textile fiber and cottonseed oil
XinFLM lacked spherical protrusions at the day before anthesis (C1, D1), but the whole ovule and flanking surface was covered in many spherical protrusions at 0 days post anthesis (DPA) (C2, D2) and 1 DPA (C3, D3), and elongated lint fibers were present at 1 DPA
Differences in gene expression between XinWX and XinFLM were most pronounced at -1 DPA (834 Differentially expressed genes (DEGs)) and 1 DPA (1000 DEGs), while only 357 DEGs were detected at 0 DPA
Summary
Plant materials used in this study contain Xinxiangxiaoji lintless-fuzzless and linted-fuzzless isogenic lines (XinWX and XinFLM, respectively); Upland cotton genetic standard line Texas. All plants were grown under standard field conditions at PaiLou experimental field, Nanjing Agricultural University, Jiangsu Province, China. All necessary permits for collecting XinFLM, Xz-142 and TM-1 were obtained from Nanjing Agricultural University, others from Cotton Research Institute, Chinese Academy of Agricultural Sciences, China. Flowers were hand-pollinated and tagged at 0 DPA, and cotton bolls were collected at -3, -1, 0, 1 and 3 DPA. Stems and leaves were collected from two-week-old seedlings, and ovules and fibers were excised from developing flower buds or bolls. All tissues collected were flash-frozen in liquid nitrogen and stored at –70°C
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