两种小麦的原生质体培养再生植株

报道两种基因型小麦昌乐5号和山农587胚性悬浮细胞系的建立和原生质体再生植株。小麦昌乐5号和山农587的胚性愈伤组织继代一年以后形成部分颗粒状和粉粒状结构,可用于悬浮培养。当分散好、生长快的胚性悬浮细胞系形成后,即可作为原生质体培养的材料。较长时间的悬浮培养容易使材料的胚性丧失,昌乐5号的胚性悬浮细胞系20天左右建成,其原生质体再生植株的频率为14/10~5(再生植株数/植板的原生质体数);而山农587的悬浮培养物需5个月以后才能用于原生质体培养,其原生质体再生植株的频率仅为4/10~6。     

应用亲缘系数分析四川小麦种质资源的遗传多样性

对代表四川小麦优良种质的40个小麦品种进行了亲缘系数分析，探讨遗传多样性，结果表明，约60％品种组合间的COP值为0，所有供试品种COP值变异范围为0．00－0．75，平均值仅为0．08，表明大多数品种在系谱上的遗传关系很小或无遗传关系，通过COP聚类分析将供试材料聚为6个类群，2个引进品种各自聚为一个类群，其余38个品种分别以繁6，阿勃，绵阳11号，成都光头4个优势亲本聚为不同的大类群与亚类群，分析认为，供试品种间系谱上的遗传差异较大，遗传多样性较高，文中对不同育种单位及不同时期小麦遗传多样性进行了分析。     

盐胁迫下小麦新品系89122的抗氧化酶活性和内源ABA含量变化的研究

利用花粉管通道法获得的转基因耐盐小麦新品系89122和其受体“陇春13”,在盐胁迫处理下,测定盐胁迫后89122和“陇春13”无性细胞系的抗氧化酶活性的变化和脱落酸的变化,结果表明,89122较其受体“陇春13”能维持较高的SOD,POD,CAT活性,ABA含量明显高于受体。     

盐胁迫下小麦新品系89122的抗氧化酶类活性变化的研究

利用受粉后的花粉管通道将高梁ＤＮＡ导入普通小麦陇春１３号,在其后代中出现了广泛变异,从中选育出已稳定遗传的丰产、抗逆性强的新品系８９１２２。经大田和盐池鉴定,８９１２２的耐盐性明显高于陇春１３号,用ＮａＣｌ（０,０．１,０．１５,０．２,０．２５ｍｏｌ／Ｌ）处理８９１２２和陇春１３号幼苗７ｄ,测定盐胁迫后其幼苗中抗氧化酶类活性的变化。结果发现,转基因小麦８９１２２较其受体陇春１３号能维持较高的ＳＯ     

小麦染色体和间期集缩染色质孔洞及内在结构的观察

对小麦根端分生组织进行常规电镜观察的结果表明：小麦染色体和间期集缩染色质中均存在孔洞，里面有由颗粒和纤维构成的、类似于骨架的结构，对有关染色体骨架和染色质骨架之间以及它们与核基质之间的相互关系进行了讨论。     

提高农杆菌介导小麦遗传转化效率的几个因素

利用gua基因的瞬时表达研究了农杆菌介导的小麦遗传转化,探讨了影响转化效率的几个因素．结果发现,当转化时茵液浓度的OD600为1．0、侵染时间为1h、超声波处理30s、共培养期间使用抗氧化剂以及用胚性愈伤组织作为外植体等,均可使转化效率得到明显提高、综合使用这些优化的转化条件,可使小麦的瞬时转化效率提高3倍多．     

Transgene inheritance and quality improvement by expressing novel HMW glutenin subunit （HMW-GS）genes in winter wheat

The expression vector pBPC30, which carries the high molecular weight glutenin subunit (HMW-GS) 1Dx5 and 1Dy10 genes, was transferred into hexaploid winter wheat cv. Jinghua No. 1, Jing411 and Jingdong No. 6 explants of immature embryos and immature inflorescence by particle bombardment. A large number of resistant transgenic plants were obtained under the selection of herbicide bialaphos or phosphinothricin (PPT). Confirmed transgenic plants of To generation showed successful integration of HMW-GS genes and bar gene into the wheat genome. T1 generation of transgenic plants can resist 20--150 mg/L PPT.Protein analysis of T2 seed by SDS-PAGE showed that HMW-GS 1Dx5 and 1DylO genes were well expressed in offspring seed of transgenic lines by co-expression with or substitution of endogenous 1Dx2 or 1DylO. In one transgenic line, TG3-74, a new protein band between endogenous protein subunits 7 and 8 (marked as 8*) of glutenin appeared,but endogenous subunit 8 (encoded by 1By8 gene) was absent. Analysis of gluten rheological quality on seed proteins of 102 T3 plants showed that the sedimentation value of 5 transgenic lines (44.2149.0 mL) was remarkably improved,59.6%---64.3% higher than that of wild type Jinghua No. 1 and Jingdong No. 6, similar to bread wheat Cheyenne (48.0 mL). Analysis of dough rheological properties of transgenic lines showed that the dough stable time of 5 transgenic lines range from 16 to 30 min, whereas the dough stable time of wild type was only between 3--7 min. Our research suggests that introducing novel HMW-GS genes into wheat is an efficient way to improve its bread-making quality.     

利用分子标记研究小麦遗传群体的赤霉病抗性鉴定方法

针对小麦赤霉病抗源苏麦3号构建的两个小麦重组自交系遗传群体苏麦3号／Alondra和苏麦3号／安农8455，采用单花接种、表土接种及自然发病3种不同的接种方法进行小麦赤霉病抗性接种鉴定，并根据苏麦3号赤霉病抗性主效QTL的连锁分子标记Xgwm 493和Xgwm 533．1分别对群体进行抗性连锁分析．检测结果表明，在温室单花接种所获得的鉴定数据中，标记的赤霉病抗性连锁效应最高，P值分别小于0．0001，抗性鉴定结果最为准确．研究表明，对小麦赤霉病这种由数量性状控制，受外界环境影响较大的真菌病害进行抗扩展性的遗传研究，应采用控温控湿条件下的单花滴注接种鉴定方法最为合适．     

Protective effects of nitricoxide on salt stress-inducedoxidative damage to wheat(Triticum aestivum L.) leaves

The changes of chlorophyll and malondialdehyde (MDA) contents, plasma membrane permeability confirmed that 0.1 and 1 mmol/L sodium nitroprusside (SNP), a donor of nitric oxide (NO) in vivo, could markedly alleviate the oxidative damage to wheat (Triticum aestivum L.) leaves induced by 150 and 300 mmol/L NaCl treatments, respectively. Further results proved that NO significantly enhanced the activities of superoxide dismutase (SOD) and catalase (CAT), both of which separately contributed to the delay of and H2O2 accumulation in wheat leaves under salt stress. Meanwhile, the accumulation of proline was apparently accelerated. Therefore, these results suggested that NO could strongly protect wheat leaves from oxidative damage caused by salt stress.     

Expression of lysine-rich protein gene and analysis of lysine content in transgenic wheat

Expression vector pBPC102, which carries winged bean lysine-rich protein (wblrp) gene and dihydropicolinate synthase (DHDPS) gene, was transferred into hexaploid winter wheat cv. Jinghua No.l, Jing411, You899 and Yangnongl5 explants of immature inflorescence and immature embryos by particle bombardment. More than 100 transgenic plants were obtained under the selection of s-(2-aminoethyl)-L-cysteine (AEC). Confirmed transgenic plants of To and TI generation by PCR and PCR-Southern blotting analyses showed successful integration of wblrp gene into wheat genome. Analysis of transgenic plant lines of T2 by Northern dot-blotting showed good expression of wblrp gene in offspring seed. The content of free lysine in leaves, contents of bound lysine and total proteins in seeds of T2 transgenie wheat lines were determined and analyzed. Among 34 tested transgenic lines, levels of free lysine content in leaves of 9 transgenic lines are 2~3times higher than un-trans-formed wild-type cultivars. Among 17 analyzed transgenic lines, bound lysine content of 4 transgenic lines is more than 10% higher than that of wild-type cultivars. Our research suggests that introducing wblrp gene into wheat is an effective way to improve its nutrition quality.     

小麦耐甘露醇变异细胞系的离体筛选及植株再生Ⅰ

目的 离体筛选小麦耐甘露醇变异细胞系并再生植株。方法 以耐盐系小麦850512r的幼穗和幼胚为外植体诱导胚性愈伤组织。以所得到的胚性愈伤组织为起始材料，采用一步和多步正筛选法离体筛选耐甘露醇变异细胞系。结果 成功获得小麦耐甘露醇变异细胞系，并再生植株。结论 小麦耐甘露醇变异细胞系的离体筛选并再生植株成功，证明在细胞水平上筛选并获得具有潜在耐水分胁迫特性的小麦新品系是完全可能的。     

Molecular identification of asymmetric somatic hybrids between wheat and Italian ryegrass

Sexual incompatibility between common wheat and Italian ryegrass was an obstacle for transferring useful traits from italian ryegrass to wheat. In order to use those desirable genetic resources to improve wheat and to create new cytoplasmic germplasm, the protoplasts of wheat and Italian ryegrass were successfully electrofused and the somatic hybrid plants were regenerated. Examination with 6 restriction enzymes, 13 probes including 9 mtDNA probes (H454, Pst24, B30, Pro I, 490, B342, pHJ2-7-1, B376, 7), 3 cpDNA probes (pHvc p1, pHvc p5 and pHvc p8) and one nuclear DNA probe-- pTA71 (rDNA) in total 73 enzyme/ probe combinations revealed rich polymorphism between the fusion partners. RFLP analysis indicated that approximately 93.4% of the regenerated plants were true somatic hybrids. AFLP analysis implied that the somatic hybrids were highly asymmetric. The RFLP analysis using mt- and cpDNA specific probes also demonstrated the non-coexistence of mitochondria and chloroplasts from the fusion partners in the somatic hybrid cells.     

BYDV CP基因转化小麦原生质体及转基因植株再生

用原生质体融合技术与ＰＥＧ介导的直接转基因方法相结合，使高频率分裂的小麦济南１７７悬浮细胞系原生质体与具有分化能力的小麦济南１７７胚性意伤组织原生质体融合，形成具有分裂和分化能力的融合细胞；同时，利用ＰＥＧ对原生质体的直接转基因作用，将带有抗病毒基因（ＢＹＤＶＣＹｇｅｎｅ）的质粒Ｐｐｐ１５与带有抗性选择标记基因的质粒严ＰＢｌＡｃｔＳＮ导入融合细胞中．经抗性筛选获得抗潮霉素（Ｈｍ）的阳性克隆并再生植株．对再生植株作ＰＣＲ检测表明，ＣＰ基因已整合到转化植株的基因组中并稳定表达．     

普通小麦的体细胞胚发生、发育相关蛋白质的毛细管电泳法鉴定

采用高效毛细管电泳技术对普通小麦不同类型的愈伤组织进行蛋白质电泳分析，发现不同类型的愈伤组织中含有多数相同的蛋白质和少数差异蛋白质．结合形态与结构分析，表明差异蛋白质可能与体细胞胚的发生和发育相关．与普通电泳及双向电泳相比，该技术具有快速可靠的特点，可望发展成为鉴定愈伤组织类型及体细胞胚发育时期特异蛋白质的有效方法．     

脯氨酸处理种子对小麦和玉米抗盐性的效应

用不同浓度的脯氨酸溶液处理小麦和玉米种子后,其发芽率、幼苗的干物质积累以及脯氨酸含量部比对照高,说明脯氨酸处理种子后,可以提高小麦和玉米的抗盐性。     

两种植物内生菌分离的影响因素研究

 在开展核桃树(Juglans regia L.)和飞龙斩血(Toddalia asiatica(L.)Lam.)植物内生菌多样性研究过程中,对季节、区域、树龄和采集部位等影响内生菌分离的因素进行研究.研究结果表明,在春夏之交宿主植物新陈代谢旺盛时采集的较大树龄材料,分离获得更多的内生菌种类和数量.采用5种不同的消毒剂,以不同的消毒浓度和消毒时间进行试验,探讨植物内生菌分离培养的最佳条件.试验表明,选用3.0%的H2O2溶液消毒30～120 s,0.5%的KMnO4溶液消毒30～120 s,75%的乙醇溶液消毒60～120 s,4%的漂白粉消毒30～120 s,对试材进行表面消毒,效果较好.从而为进一步开发利用药用植物内生菌提供了基础资料.     

小麦温光生态研究及其在生产中的应用进展

文章对小麦温光反应理论，温度和光照在小麦生长和发育过程中的相互关系，以及温光反应理论在生产实践中的应用新进展进行了总结，可为小麦温光生态的研究和小麦生产实践提供理论指导。     

Genetic diversity of Chinese summer soybean germplasm revealed by SSR markers

There are abundant soybean germplasm in China. In order to assess genetic diversity of Chinese summer soybean germplasm, 158 Chinese summer soybean accessions from the primary core collection of G max were used to analyze genetic variation at 67 SSR loci. A total of 460 alleles were detected, in which 414 and 419 alleles occurred in the 80 Huanghuai and the 78 Southern summer accessions, respectively. The average number of alleles per locus was 6.9 for all the summer accessions, and 6.2 for both Huanghuai and Southern summer accessions. Marker diversity (D) per locus ranged from 0.414 to 0.905 with an average of 0.735 for all the summer accessions, from 0.387 to 0.886 with an average of 0.708 for the Huanghuai summer accessions, and from 0.189 to 0.884 with an average of 0.687 for the Southern summer accessions. The Huanghuai and Southern summer germplasm were different in the specific alleles, allelic-frequencies and pairwise genetic similarities. UPGMA cluster analysis based on the similarity data clearly separated the Huanghuai from Southern summer soybean accessions, suggesting that they were different gene pools. The results indicate that Chinese Huanghuai and Southern summer soybean germplasm can be used to enlarge genetic basis for developing elite summer soybean cultivars by exchanging their germplasm.     

野生大豆DNA导入小麦及RAPD分子验证

利用花粉管通道法将野生大豆总DNA导入小麦，以期获得变异系小麦．对小麦球蛋白的SDS—PAGE，印迹表明，冬小麦T2l6#产生了新的蛋白亚基，其分子量为78kD，而有的变异品系一些蛋白亚基消失了；凯氏定氮法和氨基酸分析表明小麦后代T216#的总蛋白含量增加，其氨基酸组成，尤其是赖氨酸含量明显提高．RAPD分析结果表明，新品系基因组出现多态性，并具有供体特异性DNA带，这些变异现象表明该小麦品系为转基因后代．本实验为选育高蛋白、优质的新小麦品种提供了途径．