甘蔗B12D基因全长cDNA克隆与表达分析

张玉叶; 黄宁; 肖新换; 黄珑; 苏炜华; 许莉萍; 阙友雄

doi:10.15886/j.cnki.rdswxb.2014.02.011

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甘蔗B12D基因全长cDNA克隆与表达分析

张玉叶, 黄宁, 肖新换, 黄珑, 苏炜华, 许莉萍, 阙友雄

文章导航 > 热带生物学报 > 2014 > 5(2): 111-119

张玉叶, 黄宁, 肖新换, 黄珑, 苏炜华, 许莉萍, 阙友雄. 甘蔗B12D基因全长cDNA克隆与表达分析[J]. 热带生物学报, 2014, 5(2): 111-119. doi: 10.15886/j.cnki.rdswxb.2014.02.011

引用本文:

ZHANG Yuye, HUANG Ning, XIAO Xinhuan, HUANG Long, SU Weihua, XU Liping, QUE Youxiong. Cloning and Expression Analysis of Full cDNA of B12D Gene in Sugarcane[J]. Journal of Tropical Biology, 2014, 5(2): 111-119. doi: 10.15886/j.cnki.rdswxb.2014.02.011

Citation:

ZHANG Yuye, HUANG Ning, XIAO Xinhuan, HUANG Long, SU Weihua, XU Liping, QUE Youxiong. Cloning and Expression Analysis of Full cDNA of B12D Gene in Sugarcane[J]. Journal of Tropical Biology, 2014, 5(2): 111-119. doi: 10.15886/j.cnki.rdswxb.2014.02.011

甘蔗B12D基因全长cDNA克隆与表达分析

doi: 10.15886/j.cnki.rdswxb.2014.02.011

福建农林大学农业部福建甘蔗生物学与遗传育种重点实验室/国家甘蔗产业技术研发中心, 福建福州 350002

基金项目:

福建农林大学杰出青年基金(xjq201202)

教育部博士点基金(20103515120006)

国家自然科学基金(31340060)

详细信息

第一作者:
张玉叶(1990-),女,福建农林大学2012级硕士研究生.E-mail:2072352@qq.com

通信作者:
许莉萍,博导,研究员,博士,主要从事甘蔗遗传育种与生物技术研究.E-mail:xlpmail@126.com

阙友雄,硕导,副研究员,博士,从事甘蔗应用基因组学研究.E-mail:queyouxiong@126.com

中图分类号: S566.1

Cloning and Expression Analysis of Full cDNA of B12D Gene in Sugarcane

Key Laboratory of Sugarcane Biology and Genetic Breeding (Fujian), Ministry of Agriculture, Fujian Agriculture and Forestry University/Sugarcane Research & Development Center, China Agricultural Technology System, Fuzhou 350002, China

摘要: 对甘蔗(Saccharum officinarum L.)茎全长cDNA文库进行大规模测序,获得了B12D基因的全长cDNA序列,命名为ScB12D(GenBank Accession number:KF714497)。生物信息学分析表明,该基因全长771 bp,编码87氨基酸(ORF,104～367 bp);该基因编码的蛋白无信号肽,为亲水性非分泌型蛋白;有1个B12D家族保守结构域,二级结构为无规则卷曲;基因定位于质膜,参与能量代谢。同时,甘蔗ScB12D基因在不同物种间具有一定的保守性,在近缘植物中具有高度的保守性,与单子叶禾本科植物玉米、粟米、高粱及水稻的同源性超过90%,与双子叶甘薯和山茶的同源性在70%左右。荧光定量PCR表达分析结果表明,甘蔗ScB12D基因在根、蔗芽、茎(包括蔗皮和蔗髓)、叶片和叶鞘等组织中组成型表达,其中在叶鞘和根中表达量较高;在生物胁迫黑穗病胁迫下,该基因的表达量在所有时间段均呈下调趋势;在非生物胁迫时,该基因的表达受NaCl胁迫后表达量最高,在ABA胁迫下表达量次之,推测该基因的表达可能与甘蔗响应生物和非生物胁迫的机制有关。
- 甘蔗 /
- B12D基因 /
- 生物信息学 /
- 荧光定量PCR
Abstract: A full length cDNA sequence of sugarcane gene was obtained from sugarcane（Saccharum officinarum L.） stem full length cDNA library through large-scale sequencing and validated by the corresponding bioinformatics analysis,termed ScB12D（Genbank Accession number: KF714497）. The full length of ScB12 D gene was771 bp,with a 264 bp open reading frame（ ORF）,encoding 87 amino acids residues. The ScB12 D of sugarcane was a basic protein,which has a conserved functional domain with the main function for energy metabolism,and this protein was located in plasma membrane. The main secondary structure element was random coil.At the same time,the ScB12 D genes is conservative in different species,especially highly conservative in kindred plants. The B12 D gene in sugarcane has a homology of more than 90% with the same genes in the monocotyledonous gramineous plants of maize,millet,sorghum and rice,and around 70% with the same genes in the dicotyledonous plants of sweet potatoes and camellia. Real-time quantitative PCR（ RT-qPCR） analysis revealed that the expression of ScB12 D was higher in root and sheath than in leaf sheath,stem（contain pith and skin）,lateral buds and leaf. Moreover,after inoculation with Sporisorium scitamineum,the expression of this gene decreased in the different time points. Meanwhile,the expression of ScB12D in the NaCl stress was the highest in abiotic stresses and slightly lower under the abscisic acid stress. It can be inferred that the sugarcane B12 D gene be associated with the reaction mechanism of sugarcane to biotic and abiotic stresses.
- sugarcane /
- B12D gene /
- bioinformatics /
- real-time PCR

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甘蔗B12D基因全长cDNA克隆与表达分析

doi: 10.15886/j.cnki.rdswxb.2014.02.011

福建农林大学农业部福建甘蔗生物学与遗传育种重点实验室/国家甘蔗产业技术研发中心, 福建福州 350002

基金项目:

福建农林大学杰出青年基金(xjq201202)

教育部博士点基金(20103515120006)

国家自然科学基金(31340060)

作者简介:
张玉叶(1990-),女,福建农林大学2012级硕士研究生.E-mail:2072352@qq.com

通讯作者: 许莉萍,博导,研究员,博士,主要从事甘蔗遗传育种与生物技术研究.E-mail:xlpmail@126.com; 阙友雄,硕导,副研究员,博士,从事甘蔗应用基因组学研究.E-mail:queyouxiong@126.com

收稿日期: 2014-02-19

中图分类号: S566.1

关键词:

甘蔗 /

B12D基因 /

生物信息学 /

荧光定量PCR

摘要: 对甘蔗(Saccharum officinarum L.)茎全长cDNA文库进行大规模测序,获得了B12D基因的全长cDNA序列,命名为ScB12D(GenBank Accession number:KF714497)。生物信息学分析表明,该基因全长771 bp,编码87氨基酸(ORF,104～367 bp);该基因编码的蛋白无信号肽,为亲水性非分泌型蛋白;有1个B12D家族保守结构域,二级结构为无规则卷曲;基因定位于质膜,参与能量代谢。同时,甘蔗ScB12D基因在不同物种间具有一定的保守性,在近缘植物中具有高度的保守性,与单子叶禾本科植物玉米、粟米、高粱及水稻的同源性超过90%,与双子叶甘薯和山茶的同源性在70%左右。荧光定量PCR表达分析结果表明,甘蔗ScB12D基因在根、蔗芽、茎(包括蔗皮和蔗髓)、叶片和叶鞘等组织中组成型表达,其中在叶鞘和根中表达量较高;在生物胁迫黑穗病胁迫下,该基因的表达量在所有时间段均呈下调趋势;在非生物胁迫时,该基因的表达受NaCl胁迫后表达量最高,在ABA胁迫下表达量次之,推测该基因的表达可能与甘蔗响应生物和非生物胁迫的机制有关。

English Abstract

Cloning and Expression Analysis of Full cDNA of B12D Gene in Sugarcane

Key Laboratory of Sugarcane Biology and Genetic Breeding (Fujian), Ministry of Agriculture, Fujian Agriculture and Forestry University/Sugarcane Research & Development Center, China Agricultural Technology System, Fuzhou 350002, China

Received Date: 2014-02-19

Keywords:

Abstract: A full length cDNA sequence of sugarcane gene was obtained from sugarcane（Saccharum officinarum L.） stem full length cDNA library through large-scale sequencing and validated by the corresponding bioinformatics analysis,termed ScB12D（Genbank Accession number: KF714497）. The full length of ScB12 D gene was771 bp,with a 264 bp open reading frame（ ORF）,encoding 87 amino acids residues. The ScB12 D of sugarcane was a basic protein,which has a conserved functional domain with the main function for energy metabolism,and this protein was located in plasma membrane. The main secondary structure element was random coil.At the same time,the ScB12 D genes is conservative in different species,especially highly conservative in kindred plants. The B12 D gene in sugarcane has a homology of more than 90% with the same genes in the monocotyledonous gramineous plants of maize,millet,sorghum and rice,and around 70% with the same genes in the dicotyledonous plants of sweet potatoes and camellia. Real-time quantitative PCR（ RT-qPCR） analysis revealed that the expression of ScB12 D was higher in root and sheath than in leaf sheath,stem（contain pith and skin）,lateral buds and leaf. Moreover,after inoculation with Sporisorium scitamineum,the expression of this gene decreased in the different time points. Meanwhile,the expression of ScB12D in the NaCl stress was the highest in abiotic stresses and slightly lower under the abscisic acid stress. It can be inferred that the sugarcane B12 D gene be associated with the reaction mechanism of sugarcane to biotic and abiotic stresses.