油茶脂氢过氧化物裂解酶基因的克隆与序列分析
作者：
王建勇;谭晓风;龙洪旭;陈鸿鹏;刘凯;...
作者机构：
中南林业科技大学 经济林培育与保护教育部重点实验室 湖南长沙;中南林业科技大学 经济林栽培与育种国家林业局重点实验室,湖南长沙 410004
会议名称：
中国林学会经济林分会2012年学术年会
会议时间：
2012-11-01
会议地点：
浙江临安
会议论文集名称：
中国林学会经济林分会2012年学术年会论文集
关键词：
油茶;脂氢过氧化物裂解酶;基因克隆;序列分析
摘要：
以油茶近成熟种子为材料,根据油茶EST 文库中已知的脂氢过氧化物裂解酶EST 序列,利用3’RACE 与5’RACE 技术,克隆到脂氢过氧化物裂解酶的全长cDNA序列.该基因全长1648bp,,包含一个1476bp 开放阅读框长,编码491个氨基酸,5’与3’非编码分别为52bp、121bp.预测该蛋白相对分子量为54.7779KDa,等电点为6.77,是个叶绿体转运肽,N 端包含有22个疏水氨基酸残基组成的序列,不仅具有转运肽富含的丝氨酸,还含有大量转运肽中很少存在的脯氨酸.多序列比对发现油茶脂氢过氧化物裂解酶核苷酸序列与茶的相似性达到96％,因此将该基因命名co HPL.
语种：
中文
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油桐种胚再生体系的建立
作者：
李泽;谭晓风;张琳;龙洪旭;袁军;...
期刊：
经济林研究 ,2012年30(4):119-122 ISSN：1003-8981
作者机构：
中南林业科技大学经济林培育与保护教育部重点实验室,湖南长沙,410004;[李泽; 范晓明; 曾艳玲; 龙洪旭; 袁军; 张琳; 谭晓风] 中南林业科技大学
关键词：
油桐;种胚;再生体系;成苗率
摘要：
为了建立快速有效的油桐再生体系,以“葡萄桐”种胚为试材,研究了贮藏方式、消毒时间、培养基及培养条件对油桐种胚成苗的影响.结果表明:种子沙藏与在自然条件下贮藏对油桐种胚成苗的影响无明显差异；种胚的最佳消毒处理为0.1％的升汞浸泡3 min;将种胚接种在1/2MS+IAA 0.5 mg/L的培养基中成苗率最高,为98％;在培养基中加入6-BA易使种胚产生大量的愈伤组织,不利于油桐胚再生体系的建立；培养基的最适pH值为5.5；与遮光处理相比,光照能缩短种胚再生体系建立的周期.
语种：
中文
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油桐4个品种光合特性的日变化
作者：
李泽;谭晓风;袁军;龙洪旭;周乃富;...
作者机构：
中南林业科技大学 经济林培育与保护教育部重点实验室,湖南长沙 410004
会议名称：
中国林学会经济林分会2012年学术年会
会议时间：
2012-11-01
会议地点：
浙江临安
会议论文集名称：
中国林学会经济林分会2012年学术年会论文集
关键词：
油桐;净光合速率;光合特性;蒸腾速率;日变化
摘要：
利用Li-6400 便携式光合测定系统，测定了4个油桐品种光合指标的日变化。结果表明：(1)‘葡萄桐’、‘小米桐’、‘对年桐’的净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)日变化均呈单峰曲线，‘大米桐’的净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)的日变化呈双峰曲线；(2) 4个品种的胞间CO2 浓度(Ci)日变化呈不规则的“V”字型；(3) 净光合速率日变化平均值大小排序为：‘葡萄桐’＞‘小米桐’＞‘对年桐’＞‘大米桐’；(4) 影响油桐净光合速率值的主要生态生理指标大小依次为：蒸腾速率＞胞间CO2 浓度＞光合有效辐射＞气孔导度。
语种：
中文
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Isolation of high-quality total RNA from floral organs of pineapple guava (Feijoa sellowiana Berg.)
作者：
Zhang Riqing;Chen Shenglin;Deyi Yuan;Long Hongxu;Liao Ting
期刊：
,2010年:5028-5030
通讯作者：
Zhang, R.
作者机构：
College of Forestry, Central South University of Forestry and Technology, Changsha city, 410004, China;China Forestry Industry Association, Beijing, 100714, China
通讯机构：
College of Forestry, Central South University of Forestry and Technology, China
关键词：
Floral organs;MADs-box;Pineapple guava;RT-PCR;Total RNA
摘要：
Pineapple guava (Feijoa sellowiana Berg.) is a great commercial plant for its fruit quality and unique flavor. Some key genes expressed in floral organs play significant roles in governing the transition from the juvenile to the adult stage. To clone these genes, high-quality RNA of floral organs were required. In this study, total RNAs were isolated and compared from pineapple guava floral organs with four approaches including guanidine isothiocyanate-based method, SDS-based method, RNA purification kit-method and improved CTAB-based method. The results showed that high-quality total RNA was acquired only by the improved CTAB-based method but not by the other three methods. Integrity examination of total RNA by agarose gel electrophoresis revealed that three RNA strips (28S, 18S, 5.8S) were clear and bright. Anlysis with spectrophotometer showed that the A260/A280 absorbance ratio was 1.9037, suggesting that the RNA was not contaminated by DNA and protein. The total RNA was applied for RT-PCR and an approximately 350 bp-sized fragment was successfully amplified. Extraction, cloning and sequencing indicated that a MADS-box gene was identified. The results suggested that the total RNA isolated by the improved CTAB-based method is of high-quality that can be used in RT-PCR or RACE-PCR for cloning of important genes expressed in floral organs. ©2010 IEEE.
语种：
英文
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油茶种子总RNA提取及Cod8FAD基因的鉴定
作者：
胡姣;谭晓风;张琳;龙洪旭
期刊：
浙江林业科技 ,2010年30(2):17-21 ISSN：1001-3776
作者机构：
中南林业科技大学林学院经济林育种与栽培国家林业局重点实验室,湖南,长沙,410004;[龙洪旭; 胡姣; 张琳; 谭晓风] 中南林业科技大学
关键词：
油茶;RNA提取;改良CTAB法
摘要：
油茶种子胚乳中富含脂肪、多糖和酚类公合物,要获得高质量的RNA难度较大。本研究以油茶优良无性系湘林1号近成熟种子的胚乳为实验材料,在试剂盒的基础上应用改良的CTAB法提取总RNA。结果表明,使用改良后的CTAB法配合试剂盒提取RNA操作简单,且能够有效去除油茶种子中多糖和多酚类等次生物质,从而得到高质量的RNA。以获得的RNA为模板,设计简并引物,通过RT-PCR,成功从油茶种子中鉴定出了油脂合成的关键酶基因之一△8脂肪酸脱饱和酶基因,命名为Cod8FAD（Camellia oleifera delta 8 fatty acid desaturase）,该基因与茶树△8脂肪酸脱饱和酶基因相似性最高,为97%,与耧斗菜△8脂肪酸脱饱和酶基因相似性最低,为62%。
语种：
中文
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翅荚木组培快繁技术的研究
作者：
袁德义;邹锋;何小勇;帅波;龙洪旭
期刊：
中南林业科技大学学报 ,2010年30(6):60-63 ISSN：1673-923X
作者机构：
[邹锋; 袁德义; 龙洪旭; 帅波] 中南林业科技大学;浙江省丽水市林业研究所;[何小勇] 浙江省丽水市林业科学研究所
关键词：
翅荚木;组织培养;快繁
摘要：
以翅荚木的腋芽以及翅荚木幼苗嫩茎为外植体,进行组培快繁技术研究,结果表明:翅荚木适宜的初代培养基为MS+6-BA 1.0 mg·L~(-1)+IBA 0.05 mg·L~(-1);继代培养基以MS+6-BA 0.5 mg·L~(-1)+IBA0.1 mg·L-~(-1)+KT0.5 mg·L~(-1)培养,效果好;生根培养则以MS+IBA0.05 mg·L~(-1)+NAA0.05 mg·L~(-1)+KT0.1 mg·L~(-1)培养基的生根效果最好。
语种：
中文
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Identification of a novel S-RNase gene and S-genotypes of four pear (Pyrus pyrifolia) cultivars
作者：
Zhang Lin* ;Hu Jiao;Tan Xiaofeng;Long Hongxu;Yuan Deyi;...
作者机构：
[Tan Xiaofeng; Long Hongxu; Yuan Deyi; Zhang Lin; Hu Jiao] Cent South Univ Forestry & Technol, Coll Forestry, Forestry Minist, Key Lab Nonwood Forest Prod, Changsha 410004, Hunan, Peoples R China.;[Li Xiugen] Chinese Acad Agr Sci, Zhengzhou Fruit Res Inst, Zhengzhou 450009, Peoples R China.
会议名称：
4th International Conference on Bioinformatics and Biomedical Engineering (iCBBE)
会议时间：
JUN 18-20, 2010
会议地点：
Chengdu, PEOPLES R CHINA
会议主办单位：
[Zhang Lin;Hu Jiao;Tan Xiaofeng;Long Hongxu;Yuan Deyi] Cent South Univ Forestry & Technol, Coll Forestry, Forestry Minist, Key Lab Nonwood Forest Prod, Changsha 410004, Hunan, Peoples R China.^[Li Xiugen] Chinese Acad Agr Sci, Zhengzhou Fruit Res Inst, Zhengzhou 450009, Peoples R China.
会议论文集名称：
International Conference on Bioinformatics and Biomedical Engineering
关键词：
Pyrus pyrifolia;gametophytic self-incompatibility (GSI);S-genotype;S-RNase;bioinformatics
摘要：
Gametophytic self-incompatibility (GSI) is a genetically controlled mechanism to prevent inbreeding and promote out-crossing. Pyrus pyrifolia is a commercially important fruit tree that exhibits GSI. It is necessary to identify S-genotypes of cultivars for determination of cross-compatible combination prior to performing pear plantation and breeding programs. In this study, an important local cultivar `Huobali' extensively used as parent in pear breeding programs, and its three progenies `Mantianhong', `Hongsucui' and `Meirensu' were used for S-genotyping analysis by PCR-based molecular method. Agarose gel electrophoresis showed that two fragments were produced in each of the three progenies, while three in the `Huobali'. After restriction digestion and sequencing analysis, the S-alleles in the four cultivars were determined. The allele of 345 bp from `Huobali' was determined as a novel S-RNase allele that was tentatively denominated as S-44-RNase. RT-PCR revealed that the S-44-RNase was expressed specifically in the styles, which is consistent with the expression pattern of S-RNases. Comparison of genomic and cDNA sequences revealed an intron of 148 bp for the S-44-RNase. At the amino acid level, it shared 65% to 93% similarity with other Maloideae S-RNases. This study will be helpful in pear production and breeding programs.
语种：
英文
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中国白梨PbSFBB13-gamma基因的分子克隆与序列特征
作者：
张琳;谭晓风;胡姣;乌云塔娜;袁德义;...
期刊：
林业科学 ,2009年45(11):36-43 ISSN：1001-7488
作者机构：
[张琳; 谭晓风; 胡姣; 乌云塔娜; 袁德义; 龙洪旭] 中南林业科技大学资源与环境学院;[何小勇] 浙江省丽水市科普工作指导站;[李秀根] 中国农业科学研究院郑州果树研究所
关键词：
白梨;配子体自交不亲和性;花粉S基因;S位点F-box基因;进化树
摘要：
以5个已知S基因型的中国白梨品种为试材,设计分别对应于梨SFBB-alpha,SFBB-beta和SFBB-gamma 基因的3对引物对这5个品种进行基因组PCR扩增.结果仅引物组合PSFBG-F/PSFBG-R从‘金花'(S_(13)S_(18)),‘金花四号'((S_(13)S_(18))和‘鹅梨'((S_(13)S_(34))中扩增出一约1 300 bp的条带,经回收、克隆、测序后,鉴定其为SFBB-gamma基因,命名为PbSFBB13-gamma(Pyrus bretschneideri SFBB13-gamma),GenBank登录号为EU081892.逆转录PCR结果表明PbSFBB13-gamma仅在花粉中特异表达;序列分析表明该基因不含内含子,编码区含1 191个核苷酸,编码396个氨基酸,预测分子质量与等电点分别为45.4 ku、4.63.PbSFBB13-gamma表现出典型的花粉SFB/SLF基因的基本结构特征,即1个F-box结构域及4个可变区;在推导氨基酸水平上,其与蔷薇科SFB/SLF基因的相似性为17.8%-97.7%.试验结果充分表明PbSFBB13-gamma应为花粉S基因的候选基因.选取蔷薇科34个SFB/SLF基因的全长氨基酸序列,构建进化树研究基因间的进化关系.结果表明,34个SFB/SLF基因形成2个亚科特异的类群,但不形成种特异的类群,其进化规律与蔷薇科S-RNase基因一致,即花粉SFB/SLF基因的形成也是在亚科形成之后、种形成之前.研究结果有助于从分子水平上揭示中国白梨的自交不亲和性机制.
语种：
中文
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Cloning and Sequence Characterization of SFBB (S-Locus F-Box Brothers)-Gamma Genes in Chinese Sand Pear (Pyrus pyrifolia Nakai.)
作者：
Tan Xiaofeng* ;Zhang Lin;Long Hongxu;Hu Jiao;He Gongxiu
期刊：
2009 3RD INTERNATIONAL CONFERENCE ON BIOINFORMATICS AND BIOMEDICAL ENGINEERING, VOLS 1-11 ,2009年:453-456
通讯作者：
Tan Xiaofeng
作者机构：
[Tan Xiaofeng; Long Hongxu; Zhang Lin; Hu Jiao] Cent South Univ Forestry & Technol, Coll Forestry, Forestry Minist, Key Lab Nonwood Forest Prod, Changsha 410004, Hunan, Peoples R China.;[He Gongxiu] Cent South Univ Forestry & Technol, Coll Forestry, Changsha 410004, Hunan, Peoples R China.
通讯机构：
[Tan Xiaofeng] C;Cent South Univ Forestry & Technol, Coll Forestry, Forestry Minist, Key Lab Nonwood Forest Prod, Changsha 410004, Hunan, Peoples R China.
会议名称：
iCBBE 2009
会议时间：
2009-01-01
会议地点：
Beijing, China
会议论文集名称：
The 3rd International Conference on Bioinformatics and Biomedical Engineering (iCBBE 2009). [v.1]
关键词：
Chinese sand pear;Gametophytic self-incompatibility;S-genotype;S-haplotype;SFBB gene
摘要：
Gametophytic self-incompatibility (GSI) is a genetically controlled mechanism to prevent inbreeding and promote out-crossing. It is controlled by a single multi-allelic Slocus which contains two separate genes: one stylar S-RNase gene and one pollen S-gene. In gametophytic self-incompatible Japanese pear (Pyrus pyrifolia Nakai.), SFBB-gamma genes have been identified as good candidates for pollen S-genes. To investigate sequence characters of the possible pollen S-gene in Chinese sand pear, degenerate primers PSFBG-F and PSFBG-R were designed for cloning the SFBB-gamma homologies in eight Chinese sand pear cultivars. By genomic PCR, sequencing and sequence analysis, a total of nine distinct sequences were identified which displayed the basic structural characterization of SFB/SLF genes, i.e. an F-box motif and four variable regions. Of the nine sequences, five equaled to reported Japanese pear SFBB-gamma genes (S1, S2, S4, S5 and S7) and the other four were considered to be new ones which were named SFBB12-gamma, SFBB13-gamma, SFBB15-gamma and SFBB22-gamma. The four new SFBB-gamma genes were expressed specifically in pollen grains. At the deduced amino acid level, they shared 20.4% to 97.7% similarities with other SFB/SLFs of rosaceous plants. The four SFBB-gamma genes cloned in this work will be useful for studying the interaction between SFBs and S-RNases, and clarifying the mechanism of GSI at the molecular level in Chinese pear. ©2009 IEEE.
语种：
英文
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Determination of S-Genotypes of Seven Cultivars and Identification of a Novel S-RNase Allele in Pyrus pyrifolia
作者：
张琳;谭晓风;周建;何小勇;袁德义;...
期刊：
林业科学 ,2008年44(10):42-48 ISSN：1001-7488
作者机构：
[张琳; 谭晓风; 袁德义] College of Resources and Environment, Central South University of Forestry and Technology;[周建] Botanical Garden Institute,Henan Institute of Science and Technology;[何小勇] Lishui Popular Science Station of Zhejiang Province;[胡姣; 龙洪旭] Co
关键词：
砂梨;配子体自交不亲和性;自交不亲和基因型
摘要：
砂梨是重要的经济树种,表现出典型的配子体自交不亲和性,在生产和育种上需鉴定品种的S基因犁以确定品种间的亲和性.选取7个砂梨品种为试验材料,使用梨S-RNase(S基因)通用引物进行基因组PCR扩增,产物通过1.8%的琼脂糖凝胶电泳分析.结果表明:‘楚比香'等4个品种中产生了预期的2条电泳条带,而其他3个品种都只产生1条带产物,通过6%的聚丙烯酰胺电泳对该3个品种的PCR产物进一步分析,结果产物被成功分离.将7个品种中分离到的14个条带分别回收、克隆、测序及序列分析,从中鉴别出10个具有梨S-RNase基因序列特征的S基因,其中‘政和大雪梨'中494 bp的基因片段被鉴定为新的S基因,暂命名为S43-RNase(GenBank接受号EF566873).RT-PCR试验证明S43-RNase仅在化柱中特异表达,符合S-RNase的表达特征.通过比对S43-RNase的基因组序列和cDNA序列,确定其内含子大小为294 bp.在推导氨基酸水平上,S43-RNase与苹果亚科其他S-RNase表现出65%～92%的相似性.
语种：
中文
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