【摘要】：【目的】克隆特早熟春性甘藍(lán)型油菜A基因組上的sBnFLD基因,并對其進(jìn)行表達(dá)研究,為該基因的功能及其在成花轉(zhuǎn)變中的作用研究奠定基礎(chǔ)�！痉椒ā扛鶕�(jù)GenBank中已報(bào)道的擬南芥和白菜型油菜FLD同源基因的保守序列設(shè)計(jì)引物,采用PCR和RT-PCR擴(kuò)增特早熟春性甘藍(lán)型油菜86號品系(光周期不敏感)的FLD同源基因,用qRT-PCR技術(shù)檢測sBnFLD基因在86號品系不同發(fā)育時(shí)期莖、葉和莖尖中的表達(dá)情況�！窘Y(jié)果】克隆出了sBnFLD基因,命名為sBnFLD,在GenBank中的登錄號為KR003079.1。sBnFLD基因cDNA全長2 376bp,有3個(gè)內(nèi)含子,4個(gè)外顯子,編碼791個(gè)氨基酸殘基,分子量86.5ku,等電點(diǎn)8.5;sBnFLD為非分泌蛋白和非膜蛋白;sBnFLD蛋白N端有2個(gè)保守的結(jié)構(gòu)域α螺旋結(jié)構(gòu)域(SWIRM)和NAD(P)-binding-8結(jié)構(gòu)域,該蛋白有多個(gè)α螺旋和β折疊。生物信息學(xué)分析顯示,sBnFLD蛋白與已報(bào)道的甘藍(lán)型油菜未知蛋白(CDX73929.1)和電子克隆的白菜型油菜FLD(XP_009135110.1)氨基酸序列相似性達(dá)99%,與擬南芥FLD氨基酸序列相似性達(dá)87%。qRT-PCR分析結(jié)果顯示,sBnFLD基因在油菜苗期和現(xiàn)蕾初期莖、葉、莖尖中均有表達(dá),但在蕾期莖尖中表達(dá)量最高。【結(jié)論】克隆出的sBnFLD基因?yàn)楦仕{(lán)型油菜的FLD同源基因,該基因在春性特早熟甘藍(lán)型油菜開花調(diào)控中可能起著重要的調(diào)節(jié)作用。
[Abstract]:[objective] to clone and express sBnFLD gene from A genome of especially precocious vernal Brassica napus L. [methods] Primer was designed according to the conserved sequence of FLD homologous gene of Arabidopsis thaliana and Brassica campestris reported in GenBank. PCR and RT-PCR were used to amplify the FLD homologous gene of rape 86 (photoperiod insensitive), and the sBnFLD gene was detected by qRT-PCR technique in stem of 86 strain at different developmental stages. The expression of sBnFLD gene in leaf and stem tip was cloned. The accession number of sBnFLD, in GenBank was 2376 BP of KR003079.1.sBnFLD gene, with 3 introns and 4 exons, encoding 791 amino acid residues. The molecular weight is 86.5ku.The isoelectric point 8.5 sBnFLD is a non-secretory protein and a non-membrane protein. There are two conserved domains, 偽 helix (SWIRM) and NAD (P) binding-8, which have multiple 偽 helix and 尾 folding. Bioinformatics analysis showed that the amino acid sequence similarity of BnFLD protein with unknown protein (CDX73929.1) and FLD (XP_009135110.1) sequence of Brassica napus and Arabidopsis thaliana FLD was 990.The result of 87%.qRT-PCR analysis showed that the amino acid sequence was similar to that of Arabidopsis thaliana FLD. SBnFLD gene was used in the stem of rapeseed at seedling stage and early budding stage. Both leaf and stem tip were expressed, but they were the highest in bud stage. [conclusion] the cloned sBnFLD gene was the FLD homologous gene of Brassica napus. This gene may play an important role in the regulation of flowering in spring especially early maturing Brassica napus.
【作者單位】： 青海大學(xué)生態(tài)環(huán)境工程學(xué)院;青海大學(xué)農(nóng)林科學(xué)院;
【基金】：國家自然科學(xué)基金項(xiàng)目(31360345) 青海省科技廳項(xiàng)目(2016-ZJ-787) 國家“863計(jì)劃”項(xiàng)目(2011AA10A104) 科技部科技支撐計(jì)劃項(xiàng)目(2013BAD01B05-3)
【分類號】：S565.4
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本文編號：2224110