本文選題：桂花 + 擴展蛋白��； 參考：《農(nóng)業(yè)生物技術學報》2017年08期

【摘要】：擴展蛋白(expansin,EXP)是一類引起植物細胞壁松弛的蛋白質(zhì),在花開放過程花瓣細胞擴展中發(fā)揮重要作用。為了探究桂花(Osmanthus fragrans)中擴展蛋白基因在花開放中的表達特征,本研究從已構(gòu)建的桂花品種‘堰虹桂’花芽轉(zhuǎn)錄組數(shù)據(jù)庫中篩選得到12個與植物EXP基因同源性較高的Unigene序列,以盛開期花瓣為材料克隆得到其cDNA序列。通過NCBI數(shù)據(jù)庫中的比對結(jié)果,將12個Unigene序列分別命名為OfEXPA1~OfEXPA8,OfEXPB1~OfEXPB2,OfEXLA1,OfEXLB1(Gen Bank登錄號:KY968700~KY968711)。BLASTX查找比對結(jié)合ORF Finder工具分析發(fā)現(xiàn),除了OfEXPA1、OfEXPA5和OfEXLB1基因的cDNA序列5′端有少量缺失外,另外9個EXP基因具有cDNA全長序列。序列分析結(jié)果表明,9個全長cDNA推導的蛋白氨基酸殘基數(shù)為240~274個,預測的分子量為26.06~29.7 kD,理論等電點為4.77~9.74。桂花OfEXPA1~OfEXPA8和OfEXPB1~OfEXPB2蛋白具有典型的擴展蛋白結(jié)構(gòu)特征,N端含有8個保守的半胱氨酸結(jié)構(gòu)域,C端含有4個保守的色氨酸結(jié)構(gòu)域,其中大部分EXPA和EXPB蛋白含有組氨酸功能域HFD基序序列。OfEXLA1具有特征基序CDQC,OfEXLB1特征基序尚未見報道。qRT-PCR分析結(jié)果發(fā)現(xiàn)桂花12個EXP基因在‘堰虹桂’花芽不同發(fā)育過程中均有表達,其中OfEXPA1、OfEXPA3、OfEXPA5、OfEXPA6、OfEXPA7、OfEXPA8、OfEXPB1、OfEXPB2和OfEXLB1整體表達模式較為類似,花芽發(fā)育前期(B1~B6)表達量顯著高于花瓣擴展期(B7~FF),在花瓣擴展期表達量逐漸下降;而OfEXPA2、OfEXPA4和OfEXLA1在花芽發(fā)育前期表達量較低,而隨著花瓣顯著伸展表達量急劇增加,在花瓣擴展期呈現(xiàn)較高的表達。結(jié)合系統(tǒng)進化樹分析發(fā)現(xiàn)OfEXPA4和OfEXPA2分別與唐菖蒲(Gladiolus grandiflorus)GgEXPA1和月季(Rosa chinensis)RhEXPA1親緣關系較近,且OfEXPA2和OfEXPA4這兩個基因與GgEXPA1和Rh EXPA1基因的表達模式相似,推測這2個基因為花開放過程中的關鍵EXP基因家族成員;進一步根據(jù)OfEXLA1表達特征推測其亦參與調(diào)控桂花花開放。研究結(jié)果為全面理解桂花花開放機制、擴展蛋白基因功能提供理論依據(jù)。
[Abstract]:Expansinus exp) is a kind of protein that causes cell wall relaxation in plants and plays an important role in the expansion of flower petals.In order to investigate the expression characteristics of expanded protein gene in flowering of Osmanthus fragrans, 12 Unigene sequences with high homology to plant EXP gene were screened from the flower bud transcriptional database of Osmanthus manthus Fragrans.Its cDNA sequence was cloned from petals in full bloom.The other 9 EXP genes had full length cDNA sequences.The results of sequence analysis showed that the deduced amino acid residues of 9 full-length cDNA were 240N 274, the predicted molecular weight was 26.06 ~ 29.7 KD, and the theoretical isoelectric point was 4.77 ~ 9.74.The structural characteristics of OfEXPA1~OfEXPA8 and OfEXPB1~OfEXPB2 proteins in Osmanthus Osmanthus showed that the N-terminal contained eight conserved cysteine domains and four conserved tryptophan domains.Most of the EXPA and EXPB proteins contained histidine functional domain HFD motif. OfEXLA1 had the characteristic motif CDQCnOfEXLB1. The results of qRT-PCR analysis showed that 12 EXP genes of Osmanthus osmanthus were expressed in different stages of flower bud development.However, with the increase of the expression of petal significantly, it showed a higher expression in the period of petal expansion.The phylogenetic tree analysis showed that OfEXPA4 and OfEXPA2 were closely related to Gladiolus grandiflorus)GgEXPA1 and Rosa chinensis)RhEXPA1, respectively, and the expression patterns of OfEXPA2 and OfEXPA4 were similar to those of GgEXPA1 and Rh EXPA1.We speculated that these two genes were the key members of EXP gene family in flowering process, and they were also involved in the regulation of Osmanthus fragrans blooming according to the characteristics of OfEXLA1 expression.The results provide theoretical basis for understanding the mechanism of Osmanthus fragrans blooming and expanding the function of protein gene.
【作者單位】： 浙江農(nóng)林大學風景園林與建筑學院;浙江理工大學建筑工程學院;
【基金】：浙江省自然科學基金(No.LQ16C160003) 浙江省教育廳科研項目(No.Y201533133) 浙江農(nóng)林大學科研發(fā)展基金人才啟動項目(No.2016FR031) 浙江省農(nóng)業(yè)新品種選育重大科技專項(No.2016C02056-12)
【分類號】：S685.13

【相似文獻】

相關期刊論文 前5條

1 毛彥芝;;番茄和馬鈴薯擴展蛋白研究進展[J];黑龍江八一農(nóng)墾大學學報;2013年05期

2 牛艷梅;沈文濤;盧雅薇;周鵬;;番木瓜果實擴展蛋白Cp-EXP1基因表達的熒光定量PCR分析[J];生物技術通訊;2008年01期

3 荊贊革;柳李旺;龔義勤;姜立娜;孫新菊;周艷孔;;蘿卜擴展蛋白基因RsEXPB1克隆與表達特征分析[J];分子植物育種;2009年04期

4 孟玉平;曹秋芬;魏瑋;孫海峰;;兩個棗樹擴展蛋白基因cDNA全序列的克隆及分析[J];生物技術通報;2010年02期

5 孫振營;韓葉;李秀芳;馬秋詩;饒景萍;;柿采后丙烯和1 甲基環(huán)丙烯處理對兩個擴展蛋白基因表達的影響[J];園藝學報;2014年06期

相關會議論文 前1條

1 曹秋芬;孟玉平;;棗擴展蛋白基因ZjEXP1、ZjEXP2的cDNA克隆及分析(摘要)[A];第二屆全國果樹分子生物學學術研討會論文集[C];2009年

相關博士學位論文 前1條

1 徐倩;草坪草耐熱及葉片伸長相關擴展蛋白基因的克隆和功能研究[D];北京林業(yè)大學;2016年

相關碩士學位論文 前1條

1 朱旗;翦股穎PenA-4中AsEXP1基因啟動子的克隆和功能分析[D];北京林業(yè)大學;2010年

，

本文編號：1731490