火把梨14-3-3基因的克隆与序列分析

依据火把梨(Pyrus pyrifolia Nakai)14-3-3蛋白的EST序列设计基因特异引物,采用快速扩增cDNA末端技术(Rapid amplification of cDNA ends,RACE),从云南火把梨中克隆一个新的14-3-3基因(Pp14-3-3)的全长cDNA序列。Pp14-3-3全长cDNA为1 107 bp,具有84 bp 5’非编码区(Untranslated Regions,UTR)、786 bp开放读码框(Openreading frame,ORF),以及237 bp 3’UTR,编码含261个氨基酸的蛋白质。Pp14-3-3与其他物种14-3-3蛋白在中间功能区域高度保守,包含典型的14-3-3结构域。与已知植物14-3-3s家族成员间的氨基酸序列进行聚类分析,将Pp14-3-3聚为非ε大类14-3-3。Pp14-3-3在火把梨接受光照和没有光照的果皮以及幼嫩叶片中大量表达,且表达量稳定。对Pp14-3-3的基因克隆以及表达特性进行了分析,为后期深入研究该新基因的功能奠定了基础。     

翠竹及菲白竹LEA3同源基因的克隆及序列分析

竹类植物在中国有着非常悠久的栽培历史,并被广泛应用于园林、造林、庭院以及食品等行业。本研究以翠竹和菲白竹为材料提取其叶片总DNA,采用PCR方法获得翠竹SpLEA3基因与菲白竹SfLEA3-1、SfLEA3-2基因;其中SpLEA3全长804 bp,编码195个氨基酸,GC含量为68.4%;SfLEA3-1全长803 bp,编码195个氨基酸,GC含量为68.2%;SfLEA3-2全长557 bp,编码144个氨基酸,GC含量为67.8%。通过ProtParamy等生物软件分析,SpLEA3、SfLEA3-1和SfLEA3-2与贵州悬竹LEA3基因同源性高达61%以上,且3个基因编码蛋白均属于亲水性蛋白。源自2个竹种LEA3基因均包含一个完整的开放阅读框,其编码的氨基酸含有4~6个由11个氨基酸组成的保守基元序列。本研究不仅为深入了解竹类植物抗旱的分子机理研究提供了基础数据,也为竹类植物的抗旱育种后续研究提供了科学依据。     

西藏青稞LEA3蛋白新抗旱基因的克隆与序列分析

以强抗旱青稞冬青8号和弱抗旱青稞品比14为材料,将幼苗经干旱诱导处理12 h提取总RNA,RT-PCR同源克隆到2个有差异的LEA3蛋白抗旱基因的全编码框序列。冬青8号、品比14的LEA3蛋白基因序列全长分别为661 bp和694 bp,其中,来源于品比14的LEA3蛋白基因编码框全长639 bp,编码含213个氨基酸残基的蛋白质,该多肽含有9个重复的、由11个氨基酸残基组成的保守基元序列。而来源冬青8号的LEA3蛋白基因与之相比较除缺少33 bp核苷酸外,另有6个碱基的差异,所编码的蛋白质也相应地缺少第4个保守基元序列,由8个重复的保守基元序列组成。二者在DNA与氨基酸序列上的同源性分别为94.38%和92.02%。结果证实抗旱性不同的青稞品种之间LEA3抗旱蛋白保守基元拷贝数有差异,推测抗旱蛋白结构的差异可能对植物的抗旱性有影响。     

高羊茅光周期调控基因CONSTANS的克隆与分析

本研究以坪用型"黔草1号"高羊茅为实验材料,基于多年生黑麦草CONSTANS(CO)基因序列设计引物,利用3'RACE和5'RACE方法扩增CO基因。结果表明,从高羊茅中分离出CO基因,命名为FaCON-STANS。经序列分析,cDNA全长1557bp,编码376个氨基酸,具有完整的开放阅读框(ORF,166～1296bp),氨基端具有B-box型锌指结构域,碳端具有CCT(CO,CO-like,TOC1)结构域。进一步的同源性分析表明,Fa-CONSTANS蛋白与禾本科植物的CO蛋白亲缘关系较近,与其它植物的亲缘关系较远。     

峨眉拟单性木兰B类基因的克隆及系统进化分析

MADS-box基因是一类在花发育进程中起重要作用的基因。本研究通过同源克隆技术从峨眉拟单性木兰花芽中分离得到3个B-class MADS-Box基因的CDS全长,分别命名为PaomAP3、PaomPI1、PaomPI2,并对其进行生物信息学分析。结果表明,PaomAP3基因ORF(开放阅读框)长度为642 bp,编码214个氨基酸;PaomPI1基因ORF长度为633 bp,编码210个氨基酸;PaomPI2基因ORF长度为636 bp,编码211个氨基酸。氨基酸序列分析结果表明,三个基因均含有典型的MADS结构域及K结构域。PaomPI1、PaomPI2基因末端有典型的PI-derived保守基元,PaomAP3基因末端有Paleo AP3保守基元和PI-derived保守基元。木兰科植物在科内分类上存在很大争议,因此本研究利用MEGA6软件中最大简约法(MP)构建了部分木兰科植物B类基因的系统发育树,结果表明玉兰亚属与木兰亚属亲缘关系较远;拟单性木兰属独成一属,其与含笑属、玉兰亚属亲缘关系较近,而与木兰亚属亲缘关系较远。     

杉木CL14-3-3-e基因克隆及其在干旱胁迫下表达分析

14-3-3蛋白是具有组织特异性的多基因家族蛋白,与植物的抗逆功能密切相关。本研究以杉木优良无性系为材料,通过RACE技术克隆得到一条新的杉木14-3-3基因,命名为CL14-3-3-e。该基因全长1 440 bp,编码258个氨基酸,在线预测的蛋白分子量为29.70 kD,等电点为4.87,是一种酸性不稳定的疏水性非跨膜蛋白,二级结构以α螺旋为主。聚类分析发现它与地钱和小立碗藓较为亲近,进化上较为保守。荧光定量PCR检测结果显示,CL14-3-3-e基因在PEG-6000模拟的不同程度干旱胁迫下均受到不同程度诱导,且轻度干旱胁迫诱导程度要强于重度干旱胁迫,说明CL14-3-3-e参与杉木干旱胁迫响应。本研究对杉木14-3-3家族的第5个基因CL14-3-3-e进行了克隆鉴定和表达分析,为解析杉木抗旱的作用机制提供帮助。     

毛果杨PtATAF1-1转录因子基因的克隆及功能分析

ATAF1蛋白是NAC转录因子家族的成员,在植物发育过程以及胁迫应答中有着重要的调控作用。在本研究中,利用cDNA末端的快速扩增技术,从毛果杨中克隆了一个ATAF1同源基因的全长cDNA,并将相应的基因命名为PtATAF1-1。生物信息学分析表明,Pt ATAF1-1全长cDNA序列为1 242 bp,由3个外显子和2个内含子组成,编码区长873 bp。氨基酸序列多重比对分析表明,PtATAF1-1蛋白质中具有NAC家族保守的NAM结构域;杨树原生质体瞬时表达Pt ATAF1-1显示其定位在细胞核中。此外,在细菌鞭毛蛋白N端保守区域(Flg22)和脱落酸(ABA)处理下,PtATAF1-1基因的表达水平诱导发生变化。本研究对PtATAF1-1基因的同源克隆与功能分析,为深入开展ATAF1基因在杨树中的功能提供了参考依据。     

棉花生长素结合蛋白ABP1基因cDNA的克隆及序列分析

生长素结合蛋白(ABP1)在植物细胞发育中起着重要作用.在ABP1蛋白的保守区设计引物,利用RT-PCR和RACE方法,首次克隆了棉花ABP1基因cDNA的3'和5'末端.测序及生物信息学分析表明,该基因cDNA全长824 bp编码190个氨基酸,它所编码的氨基酸序列包含植物ABP1蛋白家族的所有特征,被命名为GhABP1.棉花ABP1所包含的Box A,Box B和Box C区在植物ABP1家族中是高度保守的.它的C末端氨基酸残基WDE在蛋白质的折叠及ABP1在质膜上的功能活性方面都具有重要的作用.棉花ABP1的C末端带有KDEL残基区,这个KDEL区表明它被定位在内质网上.用Clustal W软件进行多重比对分析表明,棉花ABP1与拟南芥、向日葵、油菜、辣椒、甘菊的氨基酸序列同源性分别为72%,72%,71%,70%和70%.系统进化分析表明,棉花在进化上与萝卜、拟南芥和油菜聚为一类.序列分析表明此基因为生长素结合蛋白基因家族的新成员.     

小琴丝竹和凤尾竹LEA3基因的克隆及分析

中国竹类植物资源丰富,栽培历史悠久,是集生态、经济和社会价值于一身的园林植物。本研究以小琴丝竹和凤尾竹为材料提取叶片总DNA,通过PCR扩增技术克隆得到2种竹类植物LEA3基因,其中小琴丝竹LEA3基因全长810 bp,编码区序列编码188个氨基酸,GC含量为67.9%;凤尾竹LEA3基因全长分别为810 bp和834 bp,编码区序列分别编码188个氨基酸和195个氨基酸,GC含量为67.72%和68.53%。通过DNAMAN等生物软件分析发现,小琴丝竹和凤尾竹LEA3基因与其他禾本科LEA3基因同源性均在77%以上,同源性较高,且编码蛋白均属于稳定的亲水蛋白;此外,小琴丝竹和凤尾竹LEA3基因均包含一个完整的开放阅读框,其编码的氨基酸均含有6个由11个氨基酸组成的保守基元序列,本研究不仅为分析其他竹类植物的脱水耐受性机制提供基础数据,同时也为竹类植物及其他农作物抗旱育种提供了科学依据。     

TcLr35小麦β(1,3;1,4)葡聚糖苷酶基因cDNA全长的克隆及分析

根据已发表的植物β(1,3;1,4)葡聚糖苷酶基因设计引物,利用RT-PCR和RACE技术,从被小麦叶锈菌诱导的小麦抗叶锈病基因近等基因系材料TcLr35中获得一个β(1,3;1,4)葡聚糖苷酶基因cDNA全长,命名为PR34,该基因全长序列为1416bp,具有一个编码335个氨基酸的开放阅读框(ORF),其起始密码子ATG位于13bp处,终止密码子TAG位于1015bp处,3'末端有20bp的poly(A)尾,379bp的3'非翻译区。与GenBank中小麦、大麦等多个葡聚糖苷酶基因具有较高同源性;对其推断的氨基酸序列进行分析,含有Glyco_hydro_17保守结构域,为葡聚糖苷酶基因蛋白结构域。Southern杂交显示,该基因在TcLr35小麦基因组中为低拷贝。     

The relation between wheat lines derived from the F2, F3, F4 and F5 generations for grain yield and harvest index

Summary The relationships between the F2, F3, F4 and F5 generations for grain yield were determined using random, pedigreed lines derived from each generation. The lines from two crosses were grown in plots at two sites over two years. In the first year, only F2 and F3 derived lines were available, but in the second year the F2 to F5 were grown.Correlations between lines in one generation and the mean of lines derived from them in a following generation increased as the generations were advanced. Correlations between consecutive generations were higher than those between generations two or three apart. Correlations between F2 and F5 derived lines, which indicate the effectiveness of selecting F2 lines, varied from 0.10 ns to o.49^** when lines from both generations were grown in the same environment. Correlations between years of lines from the same or different generations were low and often non-significant.Harvest index was measured on the F2 and F3 derived lines at the one site in the first year. Selection for improvement of grain yield using harvest index was no more effective than selection for yield directly, when considered across years.It is conclued that, while gains in yield can be achieved by selecting for yield in early generations, a foremost consideration needs to be the influence of different sites and years on the effectiveness of selection.     

矮牵牛F1代与F2代园艺性状的差异及其利用

1简介 在层出不穷的花卉天地间,地处南美洲的阿根廷涌现出一种被形容为"酒吧歌女"的新花-矮牵牛.由于芳容与野外的牵牛花十分相似,因而被国外专家称为矮牵牛.矮牵牛又名碧冬茄,为茄科碧冬茄属植物.矮牵牛自1835年由威廉·赫伯特(William Herbert)育成以后,1849年又出现重瓣矮牵牛品种.1876年通过自然突变育成了四倍体大花矮牵牛系列.1879年很快又推出矮生小花品种.1930年育成杂种1代的矮牵牛品种.近年来已育出抗热、抗雨和抗病品种.同时,它也是我国首批的四大转基因植物之一.     

Yield of F1, F2 and F3 hybrids of rice (Oryza sativa L.)

Summary Yield and yield components of F₁ hybrids were studied in three experiments at 30×30 cm spacings and in one experiment at 15×15 cm spacings. In the 30×30 cm experiments, 10 of the 41 hybrids tested significantly outyielded their high parents. However, only 2 hybrids significantly outyielded the best cultivar: one hybrid yielded 23 % and the other 16 % more than their respective check cultivars. The four hybrids in the 15×15 cm experiment yielded only 59 to 92% as much as their high parents.In areas where rice is transplanted at relatively wide spacings, the observed levels of F₁ heterosis in selected hybrids may be sufficient to warrant production of hybrid rice, if enough hybrid seed can be produced. For direct-seeding at the high rates normal in the USA, the relatively small levels of heterosis and the difficulties of hybrid seed production preclude use of F₁ hybrid rice cultivars at present.None of 19 bulk F₂ and F₃ hybrids in two experiments yielded significantly more than its high parent. Similarly, none of the 12 mixtures included in one experiment yielded significantly more than its high parent. On the basis of yield alone, using bulk F₂ or F₃ or simple mixture populations is not merited.Contribution from the Western Region, Agricultural Research Service, U.S. Department of Agriculture, and the Department of Agronomy and Range Science, University of California, Davis, California 95616.     

棉花杂交种F1、F2纤维品质性状比较研究

2001-2003年,通过田间试验、定位取样、分株收花、分别检测了杂交棉农杂62F1、F2的主要纤维品质指标。结果分析表明,F1、F2的纤维品质性状均为分离世代,F1、F2纤维长度、比强度、整齐度、麦克隆值、伸长率等品质指标群体平均数趋于一致,群体内个体纤维指标变异均呈正态分布,但峰值和偏斜度差异明显,F1群体中品质较优的个体显著多于F2。F2纤维的整体品质比F1差。     

甜菜M14品系BvM14-UNG基因克隆及生物信息学分析

为研究Bv-UNG蛋白在植物碱基切除修复(base-excision repair,BER)途径中的功能,以甜菜M14品系为材料,根据NCBI上二倍体栽培甜菜UDG编码基因Bv-UNG序列,利用同源序列克隆法获得甜菜M14品系BvM14-UNG基因cDNA全长,并对其进行生物信息学分析.结果 表明,该基因编码374个氨...     

转基因抗虫杂交棉杂种后代及其亲本纤维品质比较研究

2000-2004年,以转基因抗虫棉农杂62、H16、H64的F1、F2、F3及其亲本为材料,检测了主要纤维品质指标.结果表明,杂种后代的纤维长度、整齐度和亲本之间无显著差异,杂种后代纤维平均伸长率与高值亲本差异显著.但在比强度和麦克隆值方面,表现出显著的杂种优势,F1杂种优势显著地高于亲本及F2和F3.纤维比强度的中亲优势农杂62 F1平均为11.84%;农杂62F1和H16F1的高亲优势分别为9.46%、14.34%.F1、F2、F3纤维长度、整齐度、麦克隆值、伸长率等品质指标群体平均数之间虽然差异很小,但F1、F2、F3群体纤维品质性状均有相似的变异个体分布.F1群体纤维品质优于F2、F3,F2群体纤维品质略优于F3.     

Response to selection for grain yield and harvest index in F2, F3 and F4 derived lines of two wheat crosses

Summary Aspects of selection for yield and harvest index were investigated by simulating selection using data from random pedigreed F₂, F₃, F₄ and F₅ derived lines from two crosses grown in plots at two sites over two years.Improvement in yield through selection was obtained when the response was measured at the same site and in the same year as the selection. Selecting the best 10 per cent of F₂ to F₄ derived lines gave F₅ derived lines that outyielded random selections by 19 to 53 per cent for one cross and 5 to 23 per cent for the second cross. These lines were 41 to 50 per cent better than the mid-parent in one cross, but were less than the mid-parent in the other cross.However, the response to selection when measured in a different year was little better than random selection. The effect of different sites also reduced the effectiveness of selection.Selection of harvest index in early generations for improvement of yield was ineffective when response was measured at the same site in the same year, or in different years.Contrary to some theoretical proposals, the same improvement in yield was obtained by selecting in early or late generations. While high yielding genotypes may be lost by delaying selection, this is counteracted by the better predictive value of late generations due to their greater homozygosity and homogeneity.     

耐高温抗稻瘟病水稻新材料14-4606与14-4625的创制

稻瘟病是威胁四川乃至全国水稻生产最重要的病害之一,高温热害是影响中国水稻产量及其稳定性最重要的气象因素,因此,选育耐高温、抗稻瘟病水稻新材料对水稻持续稳产具有重要的意义。本研究采用辐射诱变技术、分子标记辅助选择和回交相结合的方法创制含2个抗稻瘟病基因Pi-d(t)1、Pi-d2的水稻抗稻瘟病新材料(14-4606,14-4625),这2个抗病新材料经过自然病圃鉴定,叶瘟1~3级,穗颈瘟3级,而亲本辐恢838叶瘟和穗颈瘟均为9级;在恢复力、耐高温能力上与亲本辐恢838相当。这两个抗病新材料的创制成功,为进一步改良抗稻瘟病恢复系和选育抗稻瘟病、耐高温的优良杂交稻组合奠定了坚实的基础。     

斑茅杂种后代分子检测

利用RAPD随机引物和斑茅5SrRNA间隔序列(ITS)引物对斑茅与甘蔗的杂种Fl、F2代进行了分子鉴定，获得了与含有斑茅血缘有关的RAPD分子标记3个及5SrRNA间隔序列标记1个；其中，5SrRNA间隔序列标记可用于斑茅杂种后代碱法DNA模板的快速检测。     

Phenotypic distribution of barley SSD lines and doubled haploids derived from F1 and F2 hybrids

Summary The doubled haploid (DH) system and the single seed descent (SSD) technique are frequently applied in breeding of self-pollinated crops to rapidly obtain homozygous lines from heterozygous hybrids. This study presents a comparison of populations of barley DH and SSD lines in terms of traits of stem structure. The SSD and DH lines derived from F₁ and F₂ hybrids Roland x Apex were examined in a field experiment. On the basis of a comparison of means, variances and correlations between traits in the F1DH, F2DH and SSD populations the occurrence/absence of linkage between genes responsible for the analysed traits was inferred. Independent inheritance was found for 1000-grain weight and the length of particular internodes, spike length and stem wall thickness. Moreover, no linkage was found for stem wall thickness and spike length, length of internodes I, II and thickness of stem walls, stem diameter and thickness of stem walls. The results obtained for the other pairs of traits indicate the presence of linkage.