Identification of Self-Incompatibility Genotypes in Some Sand Pears (Pyrus pyrifolia Nakai) by PCR-RFLP Analysis

The identification of self-incompatibility genotype （S-genotype） will be useful for selection of pollinizers and design of crossing in cultivar improvement of sand pear. This paper reported the identification of self-incompatibility genotypes of seven Chinese and two Japanese sand pear cultivars using PCR-RFLP analysis and S-RNase sequencing. The Sgenotypes of these cultivars were determined as follows： Huali 1 S1S3, Shounan S1S3, Xizilti S1S4, Qingxiang S3S7, Sanhua S2S7, Huangmi （Imamuranatsu） S1S6, Huali 2 S3S4, Baozhuli S7S33, Cangxixueli S5S15. S-RNase alleles （S1 to S9） in sand pear could be identified effectively by PCR-RFLP analysis.

Heterosis of Double Low Self-incompatibility in Oilseed Rape(Brassica napus L.)

66 F 1 hybrids, produced by 3 double low self-incompatible lines and 22 varieties with a North Carolina II (NCII) crossing design, were tested for their heterosis in Wuhan, China during two growing seasons from 1999 - 2001. The results showed that significant differences were found between F1s and their parents for yield per plant and seed oil content. Mid-parent heterosis of these two characters ranged from 5.50% -64.11% and from 1.55% -7.44% respectively. Heterosis for seed yield per plant was greater than that of seed oil content. For yield components, heterosis of total number of siliques per plant was the highest, followed by seed number per silique and 1 000 seeds weight. Significant genotype-by-year interaction was found for seed yield per plant. Results from correlation and combining ability analysis indicated that parental effects on its F! hybrid depended on characters, seed yield per plant was affected by both additive and non-additive effects, and seed oil content was affected mainly by additive effect. When designing hybrid programme, parents might be selected by GCAs and variances of SCAs of parents for the characters affected by both additive and non-additive effects, and by the sum of GCAs of female and male parents for the characters mainly affected by additive effects.

Self-incompatibility Characteristics of 34 Apricot Cultivars in Xinjiang

[ Objective] This study aimed to explore the relationship between self-incompatibility strength and characteristics related to pollination and fertilization of different apricot varieties in Xinjiang. [ Method] The pollen amount, pollen germination rate, pollen tube growth status and fruiting setting rate by self-pollina- tion of 34 apricot cultivars in Xinjiang were determined, to analyze the self-incompatibility of different apricot cultivars. [ Result] The average pollen amount per anther of 34 apricot eultivars was 1 213.7, and the average pollen germination rate was 46.0%. There were great differences in the self-incompatlbility of different cuhivars ; most pollen tubes of the euhivars with high self-incompatibility stopped elongating at 1/3 or 1/2 part of the styles, and only a few pollen tubes of the euhivars with low self-incompatibility reached the ovary, and the normal fertilization ratio was significantly lower than that in self-compatible cultivars. [ Conclusion] Among the 34 apricot cuhivars, only 6 cuhivars were self-compatible and the others exhibited gametophyte self-incompatibility. In addition, the fruit setting rate by self-pollination was low.

Molecular Cloning,Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage(Brassica campestris ssp.chinensis)

The exocyst is a conserved protein complex,and required for vesicles tethering,fusion and polarized exocytosis.Exo70A1,the exocyst subunit,is essential for assembly of the exocyst complex.To better understand potential roles of Exo70A1 in non-heading Chinese cabbage（Brassica campestris ssp.chinensis）,we obtained the full-length cDNA of Exo70A1 gene,which consisted of 1 917 bp and encoded a protein of 638 amino acids.BlastX showed BcExo70A1 shared 94.9% identity with Brassica oleracea var.acephala（AEI26267.1）,and clustered into a same group with other homologues in B.oleracea var.acephala and Brassica napus.Subcellular localization analysis showed BcExo70A1 was localized to punctate structures in cytosol of onion epithelial cells.Results showed that BcExo70A1 was widely presented in stamens,young stems,petals,unpollinated pistils,roots and leaves of self compatible and incompatible plants.The transcripts of BcExo70A1 in non- heading Chinese cabbage declined during initial 1.5 h after incompatible pollination,while an opposite trend was presented after compatible pollination.Our study reveals that BcExo70A1 could play essential roles in plant growth and development,and is related to the rejection of self pollen in non-heading Chinese cabbage.

海南岛越南油茶种质CdS-RNase基因及其编码蛋白的多样性分析

【目的】为了分析海南岛本地越南油茶种质中CdS-RNase基因及其编码蛋白的多样性,探索CdSRNase蛋白类型与越南油茶自交坐果率之间的关系,为依据CdS-RNase蛋白类型合理搭配越南油茶田间种质和提高越南油茶坐果率,提供科学参考。【方法】选取60份海南岛本地越南油茶种质,以其花柱c DNA为模板,RT-RCR特异扩增CdS-RNase基因序列。琼脂糖凝胶电泳检测和切胶回收CdS-RNase基因特异性扩增条带,并把特异性扩增条带连接到平末端测序载体上。将测序载体转化到大肠杆菌感受态细胞中,用带有卡那霉素抗性的LB固体培养基筛选阳性单克隆,测序得到CdS-RNase基因序列。随后,60份越南油茶种质与“CV1”~“CV18”的CdS-RNase基因序列一起用于基因单体型划分、单体型聚类分析、获得CdS-RNase蛋白氨基酸序列、CdSRNase蛋白分型、CdS-RNase蛋白二级结构预测等。最后,根据越南油茶种质中Cd S-RNase蛋白类型,把越南油茶种质划分为三类:类型Ⅰ,只检测到正常CdS-RNase蛋白;类型Ⅱ,同时检测到正常CdS-RNase和突变CdSm-RNase蛋白;类型Ⅲ,只检测到CdSm-RNase蛋白。每类越南油茶种质挑选5份,开展自交坐果率统计试验。【结果】78份海南岛本地越南油茶种质中共鉴定到171个CdS-RNase基因单体型(Hap1~Hap171),编码35种正常CdS-RNase蛋白(CdS1-RNase~CdS35-RNase)和22种突变CdSm-RNase蛋白(CdSm1-RNase~CdSm22-RNase)。正常CdS-RNase蛋白和突变CdSm-RNase蛋白二级结构中α-螺旋比例差异不显著,但是突变CdSmRNase蛋白二级结构中β-转角和β-折叠的比例极显著或显著高于正常CdS-RNase蛋白,无规则卷曲比例显著低于正常CdS-RNase蛋白。类型Ⅰ越南油茶种质自交坐果率显著低于类型Ⅱ,类型Ⅱ越南油茶种质自交坐果率显著低于类型Ⅲ。【结论】海南岛本地越南油茶种质CdS-RNase基因及其编码蛋白多样性丰富,突变CdSmRNase蛋白可能通过增加其二级结构中β-转角和β-折叠的比例减少无规则卷曲比例影响其蛋白功能的发挥,突变CdSm-RNase蛋白能一定程度上提高越南油茶自交坐果率。

欧李种质资源自交不亲和S-RNase基因的克隆及序列分析

为鉴定不同欧李种质资源的S基因型,分析欧李S基因序列特点,以70份欧李种质为材料,通过PCR特异性扩增S基因,并克隆测序得到其基因序列。结果表明:利用BFP93/BFP94-1引物,在52份种质中各鉴定到2个S基因,在16份种质中各鉴定到单个S基因,在3-32-扁黄和10-32两份种质中未扩增出条带;共克隆得到120条基因,鉴定得到36种S基因型,并将其中的新基因登录到NCBI中,登录号依次为OQ124075~OQ124109。

Parameter Self - Learning of Generalized Predictive Control Using BP Neural Network

This paper describes the self—adjustment of some tuning-knobs of the generalized predictive controller(GPC).A three feedforward neural network was utilized to on line learn two key tuning-knobs of GPC,and BP algorithm was used for the training of the linking-weights of the neural network.Hence it gets rid of the difficulty of choosing these tuning-knobs manually and provides easier condition for the wide applications of GPC on industrial plants.Simulation results illustrated the effectiveness of the method.

Self - Crosslinking Behavior of Self - Emulsion Water - Based Polyure-thane Coating Agents

The relation between structures and properties of polyurethane are investigated by modern physical and chemical methods.The results obtained are as follows:the effects of the content of self-crosslinking agent on the properties of polyurethane,i.e.,dispersion stability,dynamical viscoelasticity and mechanical properties are discussed.It is found that the optimum molar ratio of epichlorohydrin and diethylenetriamine is 1:2.A mois-

Culture Self- Consciousness in Transmission of Chinese Philosophy from the Perspective of English Globalization -- Taking the Belt and Road Initiative for Example

The Belt and Road Initiative contains the aspiration of Chinese nation to pursue ideological freedom and desire of strengthening international exchanges and cooperation. This paper takes the Belt and Road Initiative for example,emphasizing the interpretation of the inheritance and development of Chinese culture so as to study the importance of Culture Self-Consciousness in Chinese Philosophy,in the context of English globalization. That is to say,this paper stresses the spirits and the stand of nation 's demonstration on pursuing peace,cooperation and sustainable development in the civilization continuity from traditional China to the contemporary China,with practical philosophic view.

Analysis on Self-compatibility of Artificially Synthesized Brassica napus Lines from Different Cross Combinations

Brassica rapa cuhivars Qinghaidahuang and Menyuan small rape were used as female parents and separately interspecifically hybridized with Zhongchijie- lan to artificially synthesize a number of Brasica napus resources. Observation of meiosis, pollen fertility and pollen tube germination and analysis of self-compati- bility index of these artificial B. napus varieties were conducted. Results showed that pollen mother cells of artificial B. napus lines from two cross combinations all had normal chromosome behavior and high pollen fertility, but the selfing seed-setting and self-compatibility index varied extremely significantly; B. napus lines from cross combination Qinghaidahuang x Zhongchijidan are highly self-compatible, while B. napus lines from cross combination Menyuan small rape x Zhongchijielan are highly self-incompatible. This study analyzed the heredity laws of self-incompatibility trait in artificially synthesized lines and discussed applica- tion prospects of artificially synthesized self-incompatible B. napus lines in rapeseed production.

6个梨品种S基因型鉴定及新基因S_(45)-RNase序列分析

梨是典型的配子体自交不亲和果树,生产上需要配置授粉树才能获得理想的产量。鉴定梨品种的S基因型能够为合理配置授粉树提供理论依据。为鉴定S基因型,设计特异性引物对6个中国原产梨品种的基因组DNA进行扩增,片段回收、克隆、测序。使用生物信息学软件对各序列进行分析,确定6个梨品种S基因型分别为:宝珠S22S42,半斤酥S5S21,张掖长把S19S22,贵德长把S19S22,满顶雪S4S15,水冬瓜S15S45。其中S45为新分离鉴定的梨S-RNase基因,GenBank登录号为:EF643634。S45和梨S26的相似性最高,但高变区有5个氨基酸差异。

梨自交不亲和新基因S_(12)-RNase的分离鉴定及序列分析

A new selfincompatibility gene was isolated and identified from Pyrus bretschneideri cultivars of Yingzhiqing and Daaoao via PCR amplification, DNA sequence analysis and cross pollination tests. DNA sequence analysis revealed that the isolated fragment displayed a high homology with S 1 ～S 11 allele, and the identity to S 1 ～S 11 allele at the deduced amino level ranged from 56% to 72%; the high degree of variances in the hypervariable （HV） region resulted from the presence of substitution, deletion and insertion of 9 to 15 amino acids. The new Sallele was named S 12 RNase and its accession number was AY250987 in GeneBank. The sizes of HV region, intron, signal peptide, C1 region, C2 region were 39 AA, 341 bp, 27 AA, 11 AA and 10 AA, respectively. The cross pollination tests were carried out using Pyrus pyrifolia cultivars that contained S 1 ～S9RNase genes as female parents, and the cultivars of Daaoao and Yingzhiqing as male parents, respectively. The results showed that all of {%P.pyrifolia%} cultivars were compatible with Daaoao and Yingzhiqing, whereas the cross pollination between Daaoao and Yingzhiqing were incompatible, further confirming that the DNA fragment isolated was a new Sgene.

梨20个品种S基因型的鉴定及新S-RNases基因的克隆

为了鉴定我国梨品种和一些野生类型个体的S基因型,应用S-RNases特异PCR扩增、克隆和测序,对其S-RNases基因核苷酸序列进行分析,鉴定了20个梨品种和野生类型个体的S基因型。起源于我国的‘奥连’(SpS32)、‘吊蛋’(SdSe)、‘沙疙瘩’(S36Sd)品种和杏叶梨的一个类型(S22Sc)个体中存在西洋梨的S-RNases基因,证明S-RNases基因分化是在东方梨种群和西方梨种群的各个种形成之前。在秋子梨‘麦梨’、‘内蒙古山梨’中发现了2个新S-RNases基因,命名为S40-、S41-RNase(DQ903313、DQ988687)。S40-和S41-RNase基因推导的部分氨基酸序列分别与苹果属S11-和S6-RNase的同源性为100%和94.4%,这表明S-RNases的存在可能在梨属和苹果属形成之前。

扁桃SLF基因和S-RNase基因的克隆及表达分析

以扁桃‘Pioneer’品种为材料,利用RT-PCR及RACE技术,克隆了1个新的SLF(SLocusF-box)基因(PdSLF1)和两个新的S-RNase基因(PdSm和PdSn)的cDNA。PdSLF1全长1331bp,编码376个氨基酸;PdSm全长826bp,编码228个氨基酸;PdSn基因全长878bp,编码227个氨基酸。与公共数据库中的序列进行相似性比较,发现这3个基因所编码的氨基酸序列与其它蔷薇科植物相应基因的氨基酸序列均具有较高的一致性,PdSLF1为70.2%～84.8%,S-RNase为59%～83.9%。PdSLF1基因在花药中专一性表达,而PdSm和PdSn基因在雌蕊中专一性表达。

梨花柱S-RNase对花粉管超微结构的影响

采用光学显微镜和透射电子显微镜研究了离体条件下不同品种梨花柱S-RNase对异花(亲和)及自花(不亲和)花粉萌发和花粉管生长及其超微结构的影响。结果表明,花柱S-RNase抑制不亲和花粉的萌发和花粉管的生长,对亲和花粉的萌发和花粉管的生长基本没有影响。花粉生长初期,亲和及不亲和花粉管超微结构相似;但培养24h以后,亲和花粉管中充满细胞质和细胞器,而不亲和花粉管中只有靠近花粉管前端有少量细胞质,细胞壁增厚,细胞壁与细胞质之间有一层胼胝质和电子透明区间隔。

新疆扁桃品种自交不亲和S-Rnases基因型的分析鉴定

【目的】利用特异性PCR扩增、DNA测序和生物信息学方法鉴定新疆扁桃品种的S-Rnases基因型,对新疆扁桃授粉品种配置、人工授粉、遗传改良以及栽培利用都具有重要的理论意义和实践价值。【方法】以新疆24个栽培扁桃品种的叶片为试材,选用目前蔷薇科通用的引物组合对扁桃叶片基因组DNA进行自交不亲和S-RNases基因特异性PCR扩增,克隆测序结果比对分析,综合进行S-Rnases基因型的鉴定。【结果】利用引物组合AmyC5R+AS1Ⅱ对24个新疆栽培扁桃品种的基因组DNA进行S-Rnases基因特异性PCR,共扩增出32条清晰可辨的片段,克隆测序后,通过DNAMAN多重序列比对,发现共包括11个核苷酸序列,大小分别为1 688 bp(Sn1)、1 404 bp(S19)、1 330 bp(S61)、1 222 bp(S24)、1 141 bp(S25)、1 087 bp(S17)、979 bp(S10)、785 bp(S63)、777 bp(S6)、690 bp(S50)、602 bp(S15)。【结论】鉴定出了新疆扁桃品种的10个S-Rnases基因型,分别为阿买提(S15S17S63)、红宝石(S15S17)、莎舟(S15S17)、鹰嘴(S10S24)、矮丰(Sn1S25)、扁石头(Sn1S25)、桃扁桃(S6Sn1)、开心果(S6S61)、双果(S63Sn1)、纸皮(S50S61)。

南疆杏品种自交不亲和S-RNase基因型的鉴定

【目的】为鉴定新疆主栽杏品种自交不亲和S-RNase基因型,【方法】以新疆27个主栽杏品种的叶片为试材,利用蔷薇科通用引物组合:EM-PC2consFD+EM-PC3consRD、PaConsⅡ-F+PaConsⅡ-R和PruC2+PruC5对其基因组DNA进行了特异等位PCR扩增,将克隆测序结果在GenBank上进行同源性比对。【结果】结果表明,27个新疆杏品种成功扩增出54个基因片段,其核苷酸序列共包括19个不同的S-RNase基因,其中4个为GenBank上登陆的已知杏属S-RNase基因,分别定名为S24(588 bp)、S14(708 bp)、S23(972 bp)、S21(971bp),15个为新的S-RNase基因,暂时分别命名为S40(572 bp,登陆号:HQ342870、S41(401 bp,登陆号:HQ342871)、S42(357 bp,登陆号:HQ342872)、S43(493bp,登陆号:HQ342873)、S44(657 bp,登陆号:HQ342874)、S45(1 294 bp,登陆号:HQ342875)、S46(830 bp,登陆号:HQ342876)、S47(575 bp,登陆号:HQ342877)、S48(594 bp)(登陆号:HQ342878)、S49(653 bp,登陆号:HQ342879)、S50(659 bp,登陆号:HQ342880)、S51(768 bp,登陆号:HQ342881)、S52(1 520 bp,登陆号:HQ342882)、‘赛买提’S65(594 bp,登陆号:JQ327151)、‘小白杏’S66(709 bp,登陆号:JQ327152)。鉴定的27个新疆杏品种的S-RNase基因型分别为:S23S14(‘晚熟胡安娜’、‘何谢克’、‘贝新纳尔’);S52S14(‘乔尔胖’、‘馒头玉吕克’);S14S47(‘克孜阿恰’、‘元旦杏’、‘辣椒杏’);S23S4(‘莎车黑叶杏’、‘库车托咏’、‘安江胡安娜’);S52S23(‘索格佳娜丽’);S52S51(‘冬杏’);S52S50(‘苏陆克’);S52S40(‘卡巴克胡安娜’);S43S41(‘贾格达玛依桑’、‘洪得克’、‘脆佳娜丽’、‘拉胡安娜’、‘克孜佳娜丽’);S45S42(‘毛拉肖’);S45S46(‘托乎提’);S45S44(‘赛来玉吕克’);S49S24(‘库尔勒托咏’、‘黄肉油杏’);S21S65(‘赛买提’);S52S66(‘小白杏’)。【结论】研究鉴定出27个新疆杏品种自交不亲和雌蕊的S基因型,为杏品种商业化栽培配置适宜授粉树提供了理论依据。

南疆扁桃品种自交不亲和S-RNase基因型鉴定

【目的】鉴定南疆扁桃品种自交不亲和S-RNase基因型,可为科学合理地配置授粉品种提供依据,有效提高南疆扁桃的产量。【方法】以南疆10个扁桃品种的叶片为试材,利用蔷薇科通用引物PaConsⅡ-F+PaConsⅡ-R、EM-PC2consFD+EM-PC3consRD对叶片基因组DNA进行S-RNase特异性扩增,将克隆测序结果在GenBank上进行同源性比对。【结果】显示10个扁桃品种均获得2条带,共20条带,包含16种不同的S基因核苷酸序列,其中4个为GenBank上登录的已知S-RNase基因序列,分别为S1(1 370 bp)、S12(2 479bp)、S14(740 bp)、S52(1 626 bp),12个为新的S-RNase基因序列,暂时分别命名为SA(780 bp)、SB(530 bp)、SC(1 261 bp)、SD(432 bp)、SE(1 266 bp)、SF(270 bp)、SG(1 263 bp)、SH(272 bp)、SI(690 bp)、SJ(1 523bp)、SK(755bp)、SL(1 486 bp)。【结论】鉴定出10个品种的S-RNase基因型。

樱桃自交不亲和S9-单元型特异的F-box基因克隆及其表达分析

目的克隆李属甜樱桃自交不亲和性花粉S-决定子基因,为今后果树配子体自交不亲和性机理研究奠定理论基础。方法根据GenBank登录的16个S-locusF-box同源基因保守区设计兼并引物,利用RT-PCR、RACE等手段,从甜樱桃品种红灯花粉cDNA中克隆到两个编码376-氨基酸多肽的全长基因。结果GenBankBlast分析显示,克隆的两个基因中一个基因编码的蛋白产物与数据库甜樱桃自交不亲和性S3-单元型特异的PaSFB3(AB096857)编码的氨基酸序列完全一致。另一个基因编码一新的PaSFB同源序列,其推测的氨基酸序列N-端同SFB3一样具有明显的F-box基序,与PaSFB1～6的一致性为76%～82%。研究显示该基因在花粉组织中特异性表达,并表现出S9-单元型特异的连锁信号。结论新基因为甜樱桃自交不亲和性花粉S-决定子候选基因PaSFB家族中一新成员,命名为PaSFB9(GenBank登录号:DQ422809),红灯自交不亲和基因型确定为S3S9。

4个枇杷品种S基因型鉴定及新基因S_(31)-RNase序列分析

【目的】鉴定‘黄密’、‘贵妃’、‘早红’和‘软条白沙’4个枇杷品种的S基因型,为其生产栽培合理选择授粉树及杂交育种亲本选择提供科学依据。【方法】以苹果S基因高度保守区设计兼并引物对4个品种的基因组DNA进行PCR扩增,片段回收、克隆及测序,分别采用Blast软件和Bioedit软件进行同源性检索和结构分析。【结果】从参试的4个品种中共分离了4个S等位基因,分别为S2、S5、S6和S31,其中S31-RNase为新分离的枇杷S-RNase基因,Gen Bank登录号为:KC131133。所克隆获得的4个枇杷S-RNase基因均克隆到4个保守区(C2、C3、RC4和C5)和1个高变区(HV),具有与苹果S基因相同的氨基酸序列结构。【结论】确定了参试4个枇杷品种S基因型分别为:‘贵妃’S2-S6、‘黄密’S2-S5、‘早红’S5-S6、‘软条白沙’S6-S31。