自交亲和苹果品种S基因型的鉴定

为了探讨苹果自交亲和的发生机理,根据保守氨基酸序列"FTQQYQ"和"anti-1/WⅠPNV"设计了苹果花柱S基因的通用引物:P1:5＇-TTTACGCAGCAATATCAG-3＇;P2:5＇-ACGTTCGGCCAAATA/CATT-3＇,根据 GeneBank登陆的花粉SLF1(DQ422810)、SLF9(AB270792)基因设计了花粉S基因的引物分别为:P SLF1(上游5＇-3＇:CTGGTGGTGTTCTTTCCTGTGTAT;下游5＇-3＇:ACTTAACTCTTCCCTCAACTCA)和P SLF9(上游5＇-3＇CAGGTGCGTGAAAGTGAAA;下游5＇-3＇:TGAGCCAAGCATAAGAACCT),以自交亲和的苹果品种早红香为试验材料,利用PCR-RFLP、克隆、测序等方法研究早红香苹果发生自交亲和的分子性状.结果表明:自交亲和的早红香苹果2条花柱S基因没有发生变异.经BLAST比较,由花粉S基因引物P SLF1 和P SLF9扩增得到的花粉S基因的DNA序列,与MdSLF2 和MdSLF9 基因DNA序列的相似性分别为94%和96%,推导氨基酸序列相似性分别为94%和97%.由P SLF1扩增得到的花粉S基因SLF1?与MdSLF1的DNA序列相似性仅为83%,且二者不一致的碱基分布于整个DNA序列中,自交亲和苹果品种早红香花粉SLF1?基因可能变异为了MdSLF2.     

利用DDRT-PCR鉴定芸芥自交不亲和系与自交亲和系的差异表达cDNA（英文）

芸芥与芸薹属植物具有亲缘关系,也是芸薹属植物的重要育种资源。自交亲和性是芸芥的一种重要变异性状。为了探明芸芥自交亲和基因的表达器官,并分离与自交亲和性有关的cDNA片段,以芸芥的一对近等基因系,自交亲和系(SC)与自交不亲和系(SI)为试材,利用差异显示RT-PCR技术分别对开花前和开花后的叶片、花药和柱头进行鉴定。结果表明,不论开花前还是开花后,芸芥自交亲和系与自交不亲和系的叶片或花药间的扩增带型是一样的,但在近等基因系的柱头间得到了差异表达条带。这证明芸芥自交亲和基因不是组成型表达,而是组织特异性表达,柱头是其唯一的表达器官。在开花前的自交亲和系中共得到了2条cDNA片段,一条小于100 bp,另一条为750~1 000 bp,在开花后的自交亲和系中得到了1条300 bp的cDNA片段。据分析这3条cDNA片段可能与芸芥的自交亲和性密切相关。在开花前的自交不亲和系中共得到了2条cDNA片段,一条为500 bp,另一条为750 bp,同时在开花后的自交不亲和系中得到了1条500~600 bp的cDNA片段。这些cDNA片段可以用来区分芸芥的自交亲和系与自交不亲和系,还可用于克隆自交亲和基因。     

油茶CoSAUR32和CoSAUR50基因的克隆及表达分析

本研究以油茶‘华硕’为试验材料,通过PCR克隆获得两个生长素响应SAUR (Small auxin upregulated RNA)基因家族的基因,命名为CoSAUR32和CoSAUR50。CoSAUR32基因开放阅读框(ORF)为372 bp,编码123个氨基酸;CoSAUR50基因开放阅读框(ORF)为318 bp,编码105个氨基酸。序列分析发现CoSAUR32和CoSAUR50蛋白均无信号肽与跨膜结构,在叶绿体和细胞核中均可表达。对其进行同源性比对及进化树分析发现,油茶两个SAUR与茶树SAUR亲缘关系最近。实时荧光定量PCR结果表明,CoSAUR32和CoSAUR50基因在自交授粉子房中36 h时的表达量显著高于异交子房,在自交授粉花柱中24 h时的表达量显著高于异交花柱。推测CoSAUR32和CoSAUR50可能参与油茶自交不亲和过程。本研究为挖掘SAUR基因的功能及解析油茶自交不亲和分子机制提供参考依据。     

‘库尔勒香梨’自交不亲和S-RNase等位基因全长的克隆与分析

‘库尔勒香梨’是新疆重点发展的特色果树品种之一,具有自交不亲和性。本研究以新疆轮台国家果树资源圃栽培梨品种‘库尔勒香梨’为试材,利用梨S基因的保守序列设计合成的引物:‘FTQQYQ’/‘antiIIWPNV’初步确定‘库尔勒香梨’S-RNase等位基因型后,设计等位基因特异性引物,利用RT-PCR克隆两个花柱S-RNase等位基因cDNA片段,RACE(cDNA末端快速扩增)技术进行cDNA全长克隆,采用BLAST进行序列比对,Protparam等在线软件分析两个S-RNase等位基因的编码蛋白特性。根据NCBI上已登录的63条梨属S-RNase基因的全长序列,用DNAMAN软件对其氨基酸序列进行比对并用MEGA 4.0构建进化树。本试验从‘库尔勒香梨’中克隆得到了S_(22)-RNase(Gen Bank接受号:KX214125)/S_(28)-RNase(Gen Bank接受号:KX214124)等位基因cDNA的全长序列,为分子手段调控梨自交不亲和性状奠定了基础。S_(22)-RNase基因cDNA全长904 bp,ORF(开放阅读框)长684 bp,编码227个氨基酸;S_(28)-RNase基因cDNA全长921 bp,ORF长687 bp,编码228个氨基酸。BLASTP比对显示,这两个基因都具有保守的RNase-T2基因结构,属于RNase-T2家族。预测相对分子质量分别为25.6 k D和25.9 k D,等电点9.36和9.30,都属于亲水性蛋白。进化分析表明,‘库尔勒香梨’的S_(22)-RNase和S_(28)-RNase处在不同的两个分支上,表明两基因亲缘关系较远。     

利用 DDRT-PCR 鉴定芸芥自交不亲和系与自交亲和系的差异表达 cDNA

芸芥与芸薹属植物具有亲缘关系,也是芸薹属植物的重要育种资源。自交亲和性是芸芥的一种重要变异性状。为了探明芸芥自交亲和基因的表达器官,并分离与自交亲和性有关的 cDNA 片段,以芸芥的一对近等基因系,自交亲和系（SC）与自交不亲和系（SI）为试材,利用差异显示 RT-PCR 技术分别对开花前和开花后的叶片、花药和柱头进行鉴定。结果表明,不论开花前还是开花后,芸芥自交亲和系与自交不亲和系的叶片或花药间的扩增带型是一样的,但在近等基因系的柱头间得到了差异表达条带。这证明芸芥自交亲和基因不是组成型表达,而是组织特异性表达,柱头是其唯一的表达器官。在开花前的自交亲和系中共得到了2条 cDNA 片段,一条小于100 bp,另一条为750～1000 bp,在开花后的自交亲和系中得到了1条300 bp 的 cDNA 片段。据分析这3条 cDNA 片段可能与芸芥的自交亲和性密切相关。在开花前的自交不亲和系中共得到了2条 cDNA 片段,一条为500 bp,另一条为750 bp,同时在开花后的自交不亲和系中得到了1条500～600 bp 的 cDNA 片段。这些 cDNA 片段可以用来区分芸芥的自交亲和系与自交不亲和系,还可用于克隆自交亲和基因。     

果梅自交不亲和相关基因型(S、SFB/SLF)研究进展

果梅(Prunus mume Sieb. et Zucc.)原产于中国,为蔷薇科典型的配子体自交不亲和(GSI)品种。GSI至少由S位点的花柱S(S-RNase)基因和花粉S(SFB/SLF)基因决定。主要介绍了果梅GSI决定基因S基因和SFB/SLF基因的研究现状,并对基因型的鉴定技术AS-PCR的理论基础进行了介绍,对目前国内外已经鉴定出基因型的果梅品种进行了总结,这些将对果梅品种授粉树的合理配置和杂交育种的亲本选择具有重要的意义。     

新疆野扁桃自交不亲和基因SSK1的克隆及生物信息学分析

为了获得新疆野扁桃居群中自交不亲和花粉特异性表达的SSK1基因全长,以新疆野扁桃植物叶片及花粉为试验材料通过rt-PCR技术以及生物测序等技术,克隆得到了一个新的SSK1基因全长c DNA,Pet SSK1。Pet SSK1全长为602 bp,开放阅读框为534 bp,共编码了177个氨基酸,经生物信息学分析,Pet SSK1蛋白的分子式为C910H1421N233O295S6,相对分子质量为20538.0,理论推导半衰期为30 h,等电点为4.52,不稳定指数为41.48,平均疏水指数为-0.567,Pet SSK1蛋白属于水溶性不稳定蛋白。首次从新疆野扁桃植株中克隆得到了SSK1基因全长。通过对该基因的克隆及序列分析,为今后的表达分析以及有效利用该基因获得自交亲和植株奠定了基础。     

禾本科植物自交不亲和性及其分子生物学研究进展

禾本科植物自交不亲和性是禾本科植物中广泛存在的一种生殖隔离现象,有利于防止近交衰退、保持遗传变异。已报道自交不亲和禾本植物有着相同的自交不亲和系统,是一种特殊的配子体自交不亲和类型,和单一位点控制的自交不亲和性不同,其不亲和性由非连锁的两个复等位基因位点S和Z控制。自交不亲和反应中涉及了花粉蛋白质的磷酸化、Ca2 浓度的变化和蛋白激酶的参与,因此可能存在着复杂的信号传导级联反应,详细机制尚不清楚。目前S、Z基因的克隆和编码产物的鉴定均未有成功的报道,天蓝虉草可作为禾本自交不亲和性研究的一个模式作物,运用分离群体和比较基因组学方法构建S与Z位点的精细连锁图谱,以对自交不亲和基因进行图位克隆,是当前禾本科植物自交不亲和性研究的一个方向和热点。     

芸芥中DnaJ同源基因的克隆与表达分析

为研究芸芥自交亲和机理,采用mRNA差异显示反转录PCR(DDRT-PCR)分析技术,从芸芥自交亲和系开花前柱头中获得1条差异片段。测序及DNA序列的生物信息学分析表明,该差异片段与拟南芥DnaJ蛋白同源基因At J3有较高同源性,暂命名为esAtJ3基因。该序列含有一个长度为297 bp的完整开放阅读框,编码98个氨基酸残基,含有一个DnaJ-C保守结构域,属于植物DnaJ超家族。esAtJ3基因编码的蛋白无信号肽,是一个亲水性蛋白,该蛋白主要由不规则卷曲和α螺旋组成。实时荧光定量PCR分析表明,esAtJ3基因在芸芥SC系开花前柱头中表达量较高,初步推测该基因可能参与芸芥亲和性调控。     

油茶SnRK2基因家族的克隆及表达分析

以油茶(Camellia oleifera)主栽品种‘华硕’为试验材料,通过RT-RCR的方法克隆出油茶蔗糖非酵解相关蛋白激酶(sucrose non-fermenting-1-related protein kinase 2, SnRK2)基因家族,命名为CoSnRK2.5,CoSnRK2.8。利用不同的在线网站对CoSnRK2基因家族进行生物信息学分析,通过实时荧光定量PCR对CoSnRK2基因家族的表达模式进行研究分析。结果表明,CoSnRK2基因家族的CDS长度为1 035～1 092 bp,编码344～363个氨基酸,具有SnRK2基因家族2个特有的结构域:相对保守的N端结构域和特异性明显的C端结构域。实时荧光定量PCR结果表明,CoSnRK2.5基因在异交(‘华硕’ב华鑫’) 72 h花柱中的表达量明显高于自交,自交12 h子房中的表达量远高于异交;CoSnRK2.8基因在自交24 h花柱中的表达量远高于异交,异交72 h花柱中的表达量远高于自交;异交84 h子房中的表达量远高于自交。本研究对进一步挖掘该基因家族在油茶后期自交不亲和过程中发挥的作用提供研究依据,对油茶自交不亲和分子机制解析具有重要的意义。     

S-RNase genotyping and incompatibility group assignment by PCR and pollination experiments in Japanese plum

Most Japanese plum-type cultivars are self-incompatible and cross pollination is necessary to ensure fruit set. In this study, the S -RNase genotype and the incompatibility group of 68 Japanese plum-type cultivars were determined by PCR amplification of the S-RNase gene. The S -RNase genotype of 50 cultivars is first reported here and five new Japanese plum S -RNase alleles ( So , Sp , Sq , Sr , Ss ) were identified. The results obtained, together with information compiled from previous studies, allowed describing 12 new incompatibility groups (VIII–XIX). The self-incompatibility of several cultivars and the cross-compatibility among different incompatibility groups were verified by self- and cross-pollination experiments followed by observation of pollen tube growth. Five cultivars behaved as self-compatible, but two of them do not have the Se allele, which has been correlated with self-compatibility. Thus, additional sources of self-compatibility different from Se appear to be involved in Japanese plum self-compatibility.     

In vitro induction of tetraploids in colchicine-treated cocoyam plantlets

A self-compatible (SC) variant of a wild diploid potato species, Solanum chacoense, which is normally self-incompatible (SI), was investigated for the nature and genetics of self-compatibility. It was crossed with a SI cultivated diploid potato species, S. phureja. The F₁ progeny segregated SC vs. SI. Diallel crosses were made among 15 F₁'s. Self-compatibility was tested in a selfed family of a parental SC variant and in sib-mated and selfed families of F₁ progeny. All the data suggest that there is a single dominant gene (Sli) with sporophytic action inhibiting S gene expression in the pollen. Plants having a ‘Sli’ gene, produce pollen which is compatible to its own parent and plants with similar S genes. The ‘Sli’ gene has been maintained in a heterozygous condition through eight selfing generations (S₈) implying that dominant homozygotes might be associated with lethality. This revised version was published online in July 2006 with corrections to the Cover Date.     

The origin of the self-compatible almond 'Supernova'

The almond cultivar 'Supernova' is reported in the literature as a late-flowering self-compatible mutant, obtained by the irradiation of the early-flowering self-incompatible cultivar 'Fascionello'. Our work to investigate the molecular basis of this form of self-compatibility has called into question the origin of 'Supernova'. Test selfing was performed both on 'Supernova' and on the accession of 'Fascionello' from which it was derived –'Fascionello-Rome'. Both proved self-compatible. Amplification of S - RNase alleles, using consensus primers from the signal peptide region to the second conserved region of the S - RNase gene and primers specific for allele S _f , confirmed 'Supernova' and 'Fascionello-Rome' indeed have the same (in)compatibility genotype S ₁ S _f as the self-compatible cultivar 'Tuono'. Nine microsatellite markers were used to fingerprint the relevant accessions; 'Supernova' and 'Fascionello-Rome' were undistinguishable from 'Tuono'. Two Sicilian accessions, 'Fascionello' and 'Falso Fascionello', lacked S _f and differed at the SSR level, indicating no close relationship with 'Supernova'. Therefore we concluded that 'Fascionello-Rome' is actually the same as 'Tuono' and that 'Supernova' originates from 'Tuono' from which it derives its self-compatibility.     

粉蓝烟草(Nicotiana glauca)花柱S-糖蛋白的发达及对花粉管生长的影响

Self-incompatibility in flowering plants is controlled by the S-gene,encoding stylar S-glycoproteins. The isolation of the stylar glycoprotein which is the likely product of the S-gene in Nicotiana glauca has been described previously. In this paper we reported the expressiot of stylar S一glycopro     

Inheritance of sexual incompatibility in apricot

The study was carried out with 3- to 6-year-old apricot progenies from the breeding programmes of ARS in Fresno, California, USA and of CEBAS in Murcia, Spain. Progenies from 19 different crosses were studied to determine the inheritance of the genetic incompatibility trait. The 19 crosses included those where both parents were self-compatible or both self-incompatible, and also those where a self-compatible parent and another self-incompatible were crossed. Segregating progenies were studied either by bagging a sufficiently high number of flowers in balloon stage and determining the percentage of fruit set several weeks later or by following pollen tube growth with fluorescence microscopy. Results indicated that a monofactorial system with a multiallelic series appears to control the trait. Alleles for self-compatibility would allow the pollen tubes to grow in any style and reach the ovules. Alleles for self-incompatibility would stop the pollen tube growth if the same allele is present in the pollen grain and in the pistil.     

Identification and genetic characterization of different resistance sources to ascochyta blight within the genus Lens

The self-incompatibility mechanism in 10 local radish cultivars was investigated through seed set analysis and pollen tube growth behavior. The objective was to identify self-incompatible local radish lines for hybrid seed production. Based on a seed set ratio as an estimate for self-incompatibility, cultivars Tangail Local, Tasaki, Kuni, Aushi and Indian Aushi were identified as self-compatible. An intermediate seed ratio was observed in ‘Red Mollika’, ‘Pinky’ and ‘Red Bongi’. Moderately strong self-incompatibility was observed in ‘Red Kalpin’ and ‘Kuni White’. Pollen tube growth behavior revealed that self-incompatible lines had a relatively lower number of germinated pollen grains per stigma and pollen tubes per style than those of self-compatible lines. The two self-incompatibility test methods gave comparable results. None of the cultivars under study was found to be fully self-incompatible. This revised version was published online in July 2006 with corrections to the Cover Date.     

Differential pollen tube growth in inbred self-compatible almond genotypes

Pistil and pollen behavior during self- and cross-pollinations in 10 inbred almond seedlings with self-compatible genotypes and self-incompatible phenotypes were studied. Pollen from these inbred seedlings was examined for the pollen tube growth in self and non-self styles, while pistils from these seedlings were tested for their ability to support self and non-self pollen tube growth. Pistils and pollen of inbred genotypes were compatible with unrelated genotypes but the pistils were unable to support the growth of related pollen, which showed a slower tube growth rate. This may be a consequence of inbreeding, resulting in a silenced self-compatibility or cryptic self-incompatibility in some genetically self-compatible genotypes. This reaction would be a mechanism favoring crossing with unrelated genotypes and reducing inbreeding in future generations.