作物学报
作物學報
작물학보
ACTA AGRONOMICA SINICA
2015年
9期
1324-1332
,共9页
唐映红%陈建荣%刘芳%袁有美%郭清泉%昌洪涛
唐映紅%陳建榮%劉芳%袁有美%郭清泉%昌洪濤
당영홍%진건영%류방%원유미%곽청천%창홍도
苎麻%肉桂酰辅酶A还原酶基因%木质素%组织表达
苧痳%肉桂酰輔酶A還原酶基因%木質素%組織錶達
저마%육계선보매A환원매기인%목질소%조직표체
Boehmeria nivea%Cinnamoyl-CoA reductase gene%Lignin%Tissue expression
根据苎麻转录组测序信息,利用RACE法克隆出2个肉桂酰辅酶A还原酶基因全长cDNA序列,对其进行生物信息学分析,并利用荧光定量技术对苎麻快速生长期、成熟期和成熟后期该基因在木质部和韧皮部的表达模式进行研究。结果表明,BnCCR1全长1056 bp,编码277个氨基酸, Blast比对BnCCR1基因与白桦、蓖麻CCR基因核苷酸序列相似性均为70%,推测的氨基酸与蓖麻CCR基因氨基酸序列相似度为77%,蛋白质预测显示其N端存在一个3 Beta-HSD/Epimerase/NAD-binding-10的保守域；BnCCR2基因全长1291 bp,编码248个氨基酸, Blast比对BnCCR2基因与毛果杨CCR基因核苷酸序列相似度为74%,推测的氨基酸与蓖麻、毛果杨CCR基因氨基酸序列相似度均为81%,蛋白质预测显示其N端存在一个3 Beta-HSD/Epimerase/NAD-binding-4的保守域；BnCCR1和BnCCR2基因编码蛋白三维模型与矮牵牛 CCR 基因相似度分别达30.68%、44.77%,建模结果可靠；荧光定量结果显示BnCCR1和BnCCR2基因具有时期表达差异性,不具有组织表达差异,但不同组织表达量具有差异。推测BnCCR1和BnCCR2基因是存在于苎麻木质素代谢中的两种CCR基因。
根據苧痳轉錄組測序信息,利用RACE法剋隆齣2箇肉桂酰輔酶A還原酶基因全長cDNA序列,對其進行生物信息學分析,併利用熒光定量技術對苧痳快速生長期、成熟期和成熟後期該基因在木質部和韌皮部的錶達模式進行研究。結果錶明,BnCCR1全長1056 bp,編碼277箇氨基痠, Blast比對BnCCR1基因與白樺、蓖痳CCR基因覈苷痠序列相似性均為70%,推測的氨基痠與蓖痳CCR基因氨基痠序列相似度為77%,蛋白質預測顯示其N耑存在一箇3 Beta-HSD/Epimerase/NAD-binding-10的保守域；BnCCR2基因全長1291 bp,編碼248箇氨基痠, Blast比對BnCCR2基因與毛果楊CCR基因覈苷痠序列相似度為74%,推測的氨基痠與蓖痳、毛果楊CCR基因氨基痠序列相似度均為81%,蛋白質預測顯示其N耑存在一箇3 Beta-HSD/Epimerase/NAD-binding-4的保守域；BnCCR1和BnCCR2基因編碼蛋白三維模型與矮牽牛 CCR 基因相似度分彆達30.68%、44.77%,建模結果可靠；熒光定量結果顯示BnCCR1和BnCCR2基因具有時期錶達差異性,不具有組織錶達差異,但不同組織錶達量具有差異。推測BnCCR1和BnCCR2基因是存在于苧痳木質素代謝中的兩種CCR基因。
근거저마전록조측서신식,이용RACE법극륭출2개육계선보매A환원매기인전장cDNA서렬,대기진행생물신식학분석,병이용형광정량기술대저마쾌속생장기、성숙기화성숙후기해기인재목질부화인피부적표체모식진행연구。결과표명,BnCCR1전장1056 bp,편마277개안기산, Blast비대BnCCR1기인여백화、비마CCR기인핵감산서렬상사성균위70%,추측적안기산여비마CCR기인안기산서렬상사도위77%,단백질예측현시기N단존재일개3 Beta-HSD/Epimerase/NAD-binding-10적보수역；BnCCR2기인전장1291 bp,편마248개안기산, Blast비대BnCCR2기인여모과양CCR기인핵감산서렬상사도위74%,추측적안기산여비마、모과양CCR기인안기산서렬상사도균위81%,단백질예측현시기N단존재일개3 Beta-HSD/Epimerase/NAD-binding-4적보수역；BnCCR1화BnCCR2기인편마단백삼유모형여왜견우 CCR 기인상사도분별체30.68%、44.77%,건모결과가고；형광정량결과현시BnCCR1화BnCCR2기인구유시기표체차이성,불구유조직표체차이,단불동조직표체량구유차이。추측BnCCR1화BnCCR2기인시존재우저마목질소대사중적량충CCR기인。
Two cDNA sequences of cinnamoyl-CoA reductase genes were cloned by RACE technology base on transcriptome sequencing data ofBoehmeria nivea, and their bioinformatics were analyzed. Their expression levels in tissues of phloem and xylem were also tested respectively in rapid-growth stage, maturation stage and late maturity stage by quantitative Real-time PCR. The results showed that theBnCCR1was 1056 bp in length and encoding 277 amino acids. Blast and protein structure analysis showed that its cDNA sequence shared a homology of 70% compared withBetula platyphylla andRicinus communis CCR, and its amino acids did a homology of 77% compared with that ofRicinus communis. Protein prediction showed its protein N terminal with a conservative field of 3 Beta-HSD/Epimerase/NAD-binding-10. The coding sequence ofBnCCR2gene was 1291 bp, and could be translated into a 248 amino acids. Blast and protein structure analysis showed that its cDNA sequence had a homology of 74% compared withPopulus trichocarpa CCR, and its amino acids had a homology of 81% compared with these ofRicinus communis andPopulus trichocarpa. Protein prediction showed its protein N terminal with a conservative field of 3 Beta-HSD/ Epimerase/NAD-binding-4. Three-dimensional structures ofBnCCR1 and BnCCR2had relatively high similarity with CCR genes inpetunias, and the similarity was 30.68% and 44.77% respectively. The result of qRT-PCR indicated that the expression of both BnCCR1 andBnCCR2 was different in different periods, while not in different tissues, but the levels of expression in different tissues showed significant differences. We speculated thatBnCCR1 and BnCCR2are different CCR genes in lignin metabolism of Boehmeria nivea, and the result of this study provide a theoretical basis for further exploration of their function in the ramie lignin biosynthesis and regulation.
