玫瑰β-1,3-葡聚糖合成酶基因(RrCalS)克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of Rosa Rugosa β-1,3-glucan Synthase Gene (RrCalS)

作　　者：康希泉 亓帅 臧德奎[2] 郑林 徐宗大[2] 邢树堂[2] 于晓艳[2] KANG Xiquan;QI Shuai;ZANG Dekui;ZHENG Lin;XU Zongda;XING Shutang;YU Xiaoyan(Shengda Garden Engineering Co., Ltd. Shengli Oil field, Shandong Dongying 257000;Shandong Agricultural University, Shandong Tai'an 271000;Qingzhou Flower Industry Administration, Shandong Qingzhou 262500)

机构地区：[1]胜利油田胜大园林工程有限公司,山东东营257000 [2]山东农业大学,山东泰安271000 [3]青州市花卉产业管理局,山东青州262500

出　　处：《山东林业科技》2019年第2期8-11,22,共5页Journal of Shandong Forestry Science and Technology

基　　金：国家自然科学基金青年项目(31200524);国家自然科学基金面上项目(31870688)

摘　　要：为了探明胼胝质对玫瑰授粉亲和性的影响,本研究以‘唐红’玫瑰花柱为试材,采用RT-PCR和RACE方法获得了玫瑰β-1,3-葡聚糖合成酶基因的cDNA全长,命名为RrCalS。该基因全长5742 bp,开放阅读框5295 bp,编码1764个氨基酸。推导编码蛋白的分子量为204.7kD, PI值为9.00,在164-265位具有pfam14288结构域,在869-1642位具有pfam02364保守结构域,属于葡聚糖合成酶超家族;该蛋白属于疏水性、非分泌型蛋白,具有16个跨膜结构域,含有32个Ser磷酸化位点、21个Thr磷酸化位点、12个Tyr磷酸化位点;该蛋白的α-螺旋占49.49%,无规则卷曲占22.68%,β-转角占7.94%;该蛋白与Fragaria vesca等8种植物的Cals氨基酸序列同源性达73%以上,且它们的系统进化关系与传统分类结果一致。本研究为进一步深入研究玫瑰授粉不亲和机理,提高玫瑰育种理论和技术水平奠定了基础。In order to reveal which role the callose played in R. rugosa pollination incompatibility, the full-length cDNA sequence of β-1,3-glucan synthase gene was cloned for the first time from the stylus of Rosa rugosa "Tanghong" with RT-PCR and RACE methods and named as RrCalS. The full-length cDNA is 5742 bp with an open reading frame of 5295 bp, encoding 1764 amino acids. The derived protein has a molecular weight of 204.7 kD, a calculated pI of 9.00, a pfam14288 conserved domain at position164-265, a pfam02364 conserved domain at position 869-1642, and belongs to glucan synthase superfamily. The derived protein is a hydrophobic and nonsecretory protein. There are 16 transmem-brane domain, 32 Ser phosphorylation sites, 21 Thr phosphorylation sites, 12 Tyr phosphorylation sites. There are 49.49% α-helixes, 22.68% random coil, 7.94% β-corner struc-ture. This protein and the Cals protein from eight other species, including Prunus persica, share a sequence homology of greater than 73%. Furthermore,their phylogenetic relationships are consistent with their traditional classifications. These results were meaningful to reveal the molecular mechanism of R. rugosa pollination incompatibility and improve the theory of breeding ornamental R. rugosa.

关 键 词：玫瑰 β-1 3-葡聚糖合成酶 基因克隆 

分 类 号：S685.12[农业科学—观赏园艺]