棉花学报
棉花學報
면화학보
COTTON SCIENCE
2010年
2期
99-103
,共5页
朱一超%孙磊%宋焕%方天荣%郭旺珍%张天真
硃一超%孫磊%宋煥%方天榮%郭旺珍%張天真
주일초%손뢰%송환%방천영%곽왕진%장천진
棉花纤维%GhDIS2%克隆%酵母
棉花纖維%GhDIS2%剋隆%酵母
면화섬유%GhDIS2%극륭%효모
cotton fiber%GhDIS2%molecular cloning%fission yeast
棉花纤维由胚珠外被单个表皮细胞分化发育而成,其分化和伸长涉及细胞骨架的重排.本研究根据已报道的植物DISTORTED2基因,同源候选基因法克隆棉花GhDIS2.该基因包括975 bp ORF,编码324个氨基酸.qPCR分析表明,它在陆地棉TM-1不同来源的组织中均表达,于开花后12 d的纤维中达到表达高峰,开花后18 d表达开始下降,根和茎的表达量较低.Southem杂交结果表明,GhDIS2在四倍体棉花基因组中存在2个拷贝.克隆GhDIS2到酵母表达载体中,并转化到裂殖酵母中,诱导该基因过量表达,GhDIS2促使细胞扩大.初步推测,GhDIS2和其它植物DIS2功能相似,参与植物细胞肌动蛋白骨架的组装.
棉花纖維由胚珠外被單箇錶皮細胞分化髮育而成,其分化和伸長涉及細胞骨架的重排.本研究根據已報道的植物DISTORTED2基因,同源候選基因法剋隆棉花GhDIS2.該基因包括975 bp ORF,編碼324箇氨基痠.qPCR分析錶明,它在陸地棉TM-1不同來源的組織中均錶達,于開花後12 d的纖維中達到錶達高峰,開花後18 d錶達開始下降,根和莖的錶達量較低.Southem雜交結果錶明,GhDIS2在四倍體棉花基因組中存在2箇拷貝.剋隆GhDIS2到酵母錶達載體中,併轉化到裂殖酵母中,誘導該基因過量錶達,GhDIS2促使細胞擴大.初步推測,GhDIS2和其它植物DIS2功能相似,參與植物細胞肌動蛋白骨架的組裝.
면화섬유유배주외피단개표피세포분화발육이성,기분화화신장섭급세포골가적중배.본연구근거이보도적식물DISTORTED2기인,동원후선기인법극륭면화GhDIS2.해기인포괄975 bp ORF,편마324개안기산.qPCR분석표명,타재륙지면TM-1불동래원적조직중균표체,우개화후12 d적섬유중체도표체고봉,개화후18 d표체개시하강,근화경적표체량교저.Southem잡교결과표명,GhDIS2재사배체면화기인조중존재2개고패.극륭GhDIS2도효모표체재체중,병전화도렬식효모중,유도해기인과량표체,GhDIS2촉사세포확대.초보추측,GhDIS2화기타식물DIS2공능상사,삼여식물세포기동단백골가적조장.
Cotton fibers are single-celled seed trichomes of major economic importance. Many important genes are expressed during cotton fiber development. DISTORTED2 encodes the Arabidopsis homolog of the ARPC2 subunit of the ARP2/3 complex, which binds to the sides of existing actin filaments and efficiently nucleates new filaments. The cytoskeleton arrangement processes in the cotton fiber initial and elongation. In this study, the Gossypium homology of the ARPC2 subunit named asGhDIS2 was isolated from the TM-1 ovule on the day of flowering. GhDIS2 contained 975 bp open reading flame, encoding apolypeptide containing 324 amino acids. GhDIS2 was expressed constitutively in every tissue with different expression levels,with the peak in 12 days post anthesis (DPA) fiber cells and low expression levels in 18, 24 DPA fiber cells, roots and stems.Southern blotting showed that GhDIS2 had two copies in allotetraploid cotton genome. A shuttle vector contained GhDIS2 cDNA sequence was transformed into fission yeast. GhDIS2 led to a significant increase in cell length and width. These results indicated that GhDIS2 had the same function to DIS in other plants.