导入外源总DNA获得大豆早熟新品系

本文报道了在大豆自花授粉后，利用其形成的花粉管通道，将提取的含有早熟血缘供体大豆绥农８号的总ＤＮＡ，直接导入受体大豆黑农２６号中。在后代Ｄ２代中获得３个早熟株系，熟期比受体提早１５天，并迅速稳定。经异地鉴定，Ｄ９０－１０７２品系产量比对照品种提高１９．１％，于１９９３年进入省区试。经过对该组合的受体，供体及转化后代进行ＲＡＰＤ分析，证明供体ＤＮＡ片段进入受体引起后代基因组的变异。     

分子标记结合田间观察快速鉴定外源DNA导入玉米的变异后代

本研究通过花粉管通道导入技术将甜高粱总DNA导入优良玉米自交系4112和8902中,并应用RAPD标记技术从导入后代中筛选出变异后代。从306个RAPD引物中筛选出79个引物对供受体及导入后代进行鉴定,其中10个随机引物在供、受体和导入后代间有多态性。对D1和D2代进行田间茎秆糖分调查,并对D2代室内考种,发现糖分含量和穗长、穗重、轴粗、穗粒重等穗部性状都发生了明显变异。结合分子鉴定结果,最终获得了一批变异材料。     

水稻花粉管通道法导入高粱DNA的SSR分子验证

从60对SSR引物中筛选出17对,对供体高粱、受体水稻及其外源。DNA导入后代稳定材料进行了分子验证,结果表明供体高粱与受体水稻间存在多态性、受体水稻与后代稳定材料间出现多态性,供体高粱与部分后代稳定材料间出现相同的扩增带,部分供体与受体共有的特征带在后代中消失、而有些后代出现了供体和受体均没有的扩增带。这表明供体高粱的DNA不同片段已整合到水稻品种中,获得了不同的变异材料,这些。DNA片段能在后代中稳定遗传。     

外源野生胡麻总DNA遗传转化栽培胡麻及RAPD分子验证

为了克服亚麻属不同种间存在的生殖隔离,从而利用亚麻野生种优异基因进行资源创制,采用花粉管通道法将亚麻野生种垂果亚麻的基因组DNA导入到栽培种坝选三号中,获得T0种子。T1播种后所获24个单株中有3株发生明显变异,转化后代的株高、工艺长度、分茎数、分枝数及单株蒴果数与受体坝选三号相比较,均发生明显增加。利用18条引物对供体、受体及24株T1导入材料的基因组DNA进行了RAPD扩增分析,从中筛选到6条多态性引物(OPH20、OPK14、S103、S118、OPJ4、OPT6)对8个样品的DNA扩增产物带型出现明显差异,占全部所用引物的33. 3%。RAPD分析表明,D14、D17、D22 3个植株的RAPD谱带中均有来自于供体的特异条带,结合考种数据,认为是供体DNA片段成功整合进了受体基因组中引起了表型上的变化。利用花粉管通道法向胡麻栽培种中导入野生种基因组DNA能够在当代引起多种变异,同时也可在分子水平上检测到基因组的差异,推测大片段外源DNA同源重组可能是变异的主要原因。     

离子束法导入大豆DNA小麦的蛋白质含量分析

为了提高小麦蛋白质含量,采用离子束介导法将'豫豆23号'(Glycine max)的基因组DNA导入普通小麦(Triticum aestivum L.)品种'中育5号'和'淮阴9628',获得了114株T1代高蛋白含量(15%)单株,蛋白质含量最高达21.44%,显著提高了受体亲本小麦蛋白质含量。从T1到T3代连续单株选择,获得了16个遗传上基本稳定的T3代株系。结果表明,离子束介导大豆DNA导入小麦,可显著提高转化后代株系的蛋白质含量。随着世代的提高群体蛋白含量有逐步提高的趋势,在T3代基本形成遗传稳定的高蛋白品系。该方法是小麦蛋白质含量改良的可行途径。     

秋水仙素诱导小豆性状变异研究

本研究用0.2%和0.4%的秋水仙素溶液浸泡小豆京农6号(JN6)种子12 h,M_1成株率分别为26.7%和3.5%,变异率为1.24%和1.59%,结果表明处理浓度越大,成株率越小、变异率越大.0.2%秋水仙素处理12 h为较佳处理,但0.4%处理的变异类型相对丰富.M_2代与对照相比,单株粒数在5%水平上达到显著差异,百粒重在1%水平达到极显著差异;M_3代百粒重与对照相比在1%水平达到极显著差异,诱变后代中筛选出了3个高产优良株系.在M_3代共筛选到263个叶色、叶形、蔓生、多分枝、黑荚、浅红和深红粒色、大粒及早晚熟等性状变异突变体.叶形有剑叶、小密叶、肾叶突变类型;叶色有深绿、浅绿、黄化、黄斑叶变异类型.早熟和晚熟变异株分别比对照早熟10～15 d和晚熟7～10 d.     

大麦DNA导入小麦诱导抗白粉病变异的研究

本试验利用外源DNA导入技术,研究了大麦DNA导入小麦品种后,D_1,D_2和D_3代的抗白粉病变异株(系)在不同条件下的抗性表现及产量构成、籽粒蛋白质和氨基酸含量的变化。试验结果表明,导入外源DNA的D_1代小麦檀株出。现了抗白粉病等多种变异类型,其中免疫和高抗白粉病变异株古2.77％,且抗性能够向子代传递,其D_2代在大田自然发病和温室接菌条件下,有5个株系抗性保持稳定,8个株系有分离,其中一个株系在D_2和D_3代抗性均稳定。在田间D_2代有2个稳定株系(D_2-20,D_2-29)的籽粒粗蛋白质含量,比受体分别高20.3％和15.76％,17种氨基酸总量分别高23.4％和27.5％。在温室这些性,状的数值也明显高于受体。     

λDNA导入中国春小麦引起形态学及细胞学变化

本研究将λDNA导入中国春小麦，对其后代D1，D2代的花粉母细胞(PMC)减数分裂行为进行了细胞学观察，并对形态性状的变异进行了跟踪统计。D1、D2代的减数分裂过程中均出现了单价体、染色体落后、染色体桥、多极分裂等异常现象。D1代PMC减数分裂变异率为25．54％，但未发现明显的形态性状变异。D2代PMC减数分裂变异率为24．05％，与D1代相比，D2代的PMC减数分裂变异开始分化，观察的出现PMC减数分裂变异的30个单株中变异幅度为9．04％～57．14％。D2代群体形态性状发生了丰富的变异，在株高、茎秆硬度、穗长、穗粒数等方面出现了变异，并且出现了茎秆坚硬、穗长增长、穗粒数增多的优良个体。     

大豆黑农33DNA导入水稻恢复系R73的研究

用大豆黑农３３做供体，水稻恢复系Ｒ７３做受体，化粉管通道法进行外源ＤＮＡ导入。共处理颖花１５１朵，当代结实粒５２粒，Ｄ１代成苗２１株，获４株变异株，变异转化率７．７％。变异后代在抽期其株高，穗长、穗型、千粒重、每产粒娄航籽粒白质含量等性状均产生明显变异，经过儿代选择，Ｄ４代有２个株系的米质优于受体恢７３，其闰透明度高，垩白率低，具有很大的选择前途。     

闽引圆叶决明两个辐射变异新品系的若干特性研究

利用 60Coγ射线辐射闽引圆叶决明86134牧草种子,通过M1-M3代田间试验观察和筛选,筛选到两个86134的辐射变异后代,进行M4、M5代田间小区品比试验,结果表明：2个变异品系的茎叶比大幅度下降,其生物量分别增长39.26%和38.04%,达极显著水平,86134-32-3的粗脂肪含量极显著增长。RAPD分析表明,辐射后代与原种的DNA指纹图谱存在明显差异。     

外源DNA导入小麦后的变异系生物学特性及胚乳蛋白的研究

应用受粉后的花粉管能通道Ｃ４作物高梁ＤＮＡ和抗逆性强的长穗偃麦草ＤＮＡ导入小麦，结果在不同组合中出现了不同类型的广泛变异，在三个组合中已选育出几个稳定遗传的优良变异系，其生物学性状大多是介于原受体和供体之间。主要特性是叶功能期延长，籽粒千粒重和单株粒重增加，增产显著，抗锈病能力增强等，其胚乳蛋白电泳图谱发季了明显变化，在变异系中产生了新的蛋白质组分，而且Ａ区和Ｂ区增加的是蛋白质群，相反原受体６８－     

外源DNA直接导入甜玉米自交系后代性状变异

提取糯质玉米DNA利用花粉管通道法导入甜玉米自交系，其后代植株产生了变异，主要表现在株高、穗位高、叶面积等性状上的差异。经三代种植观察，D3代各变异性状趋向稳定。经品质分析，可溶性糖含量基本保持不变，蛋白质，淀粉含量有所提高。从而为今后玉米不同种之间的DNA导入创造新的变异品系，提供可参考的理论依据。     

利用回交法快速选育高油酸花生新品系

以普通花生品种花育22为母本、高油酸花生品种开农176为父本杂交得到F1杂种,筛选油酸含量高于60%且同时含有FAD2a和FAD2b位点的F1为杂交父本,以花育22为轮回亲本(母本)连续回交得到BC1F1~BC4F1代回交种。利用近红外光谱仪测定F1及BC1F1~BC4F1籽粒的油酸、亚油酸含量,选择油酸含量大于60%的种子,用刀片切取种子小部分子叶提取DNA,以F0.7/R3为引物进行PCR扩增及测序,根据测序峰图差异表现筛选出同时含有FAD2a和FAD2b位点的种子作为下一代回交的父本。切去部分子叶的种子切口用石蜡封闭,播种前浸泡于40℃温水中催芽,对12 h后未露白的种子用100 mg L–1乙烯利浸泡4 h后再转入40℃温水浸泡至24 h,发芽率可达到98%。2013年春季开始杂交,2016年春在青岛播种BC4F2代种子,取幼苗期幼叶鉴定基因型,筛选出基因型为aabb的单株,收获时选留农艺性状类似于花育22的优良单株,再利用近红外光谱仪测定所选单株油酸含量,获得油酸含量在70%以上、油酸亚油酸比值大于7.0的单株24个。这些单株与花育22相比,农艺性状基本相同,称为改良花育22高油酸花生新品系。     

Associations among widely used French and U.S. maize hybrids as revealed by restriction fragment length polymorphisms

Summary Eighty DNA Restriction Fragment Length Polymorphism (RFLP) clones were used as probes to profile 47 hybrids of maize (Zea mays L.) that are of widespread usage in France and 49 hybrids that are either in common usage or are new releases in the U.S. The objectives were to 1) investigate the degree to which RFLPs provide unique characterization of hybrids; 2) show associations among hybrids using both cluster and principal coordinate analyses; 3) measure the ability of RFLPs to show associations among hybrids that reflect those to be expected on the basis of pedigree; and 4) compare the patterns and extent of genetic diversity among French hybrids with that found among a set of widely used U.S. hybrids.RFLPs showed all French hybrids to have different profiles, however, 3 hybrids were very similar with more than 90% of their profiles in common. Twenty-seven U.S. hybrids showed this level of similarity with one or more U.S. hybrids. High correlations (r=0.93, 0.94) were found for pedigree distance versus RFLP distance between pairs of French and of French and U.S. hybrids, respectively. Similar levels of correspondence for rank correlations between RFLP and pedigree data were also found. Similar groupings of hybrids were shown by two cluster analysis methods and by principal coordinate analysis. Inclusion of hybrids in cluster groupings was supported by observation of raw distance data for selected hybrids and their nearest neighbors. Most hectarage in France is planted to hybrids that fall within 2 related groups of germplasm on the basis of RFLP data. Minimum distance standards could promote breeders to surmount the challenge of introducing elite yet diverse germplasm into agriculture.     

利用MAS技术改良香稻‘R15’的直链淀粉含量

R15是适应性较广,产量较高的中熟晚籼香稻品系,但由于含有Wx^a基因,其直链淀粉含量高达23.5%,严重影响食用口感。因此,我们以含有Wx^b等位基因的不育系Y58S为供体,通过杂交、回交,利用RAD测序技术、分子标记辅助筛选等技术,经过8个世代的筛选,成功用Wx^b基因替换掉R15中的Wx^a基因,培育出1个新的纯合株系——‘深华R16’,其染色体上有10882234 bp长度的碱基,基因型和供体亲本‘Y58S’一致,占2.92%;有362363285 bp长度的DNA片段基因型和受体亲本‘R15’一致,占97.08%;其直链淀粉含量为14%。将‘Y58S’分别与‘深华R16’和‘R15’进行组配,结果表明,‘Y58S’/‘深华R16’组合的直链淀粉含量(13.9%)低于‘Y58S’/‘R15’组合的直链淀粉含量(19.9%),其它米质性状与农艺性状基本一致。本研究为分子标记辅助育种提供了新思路,培育出一新的水稻品系。     

Genetic diversity in soybean as determined by RAPD and microsatellite analysis

The random amplified polymorphic DNA (RAPD) and microsatellite techniques were used to evaluate the genetic diversity among 18 soybean genotypes selected for a breeding programme to increase the protein content of varieties adapted for central European growing conditions. Out of 33 random primers used in RAPD reactions, only 12 showed polymorphism useful for characterization of these genotypes. In contrast, all 12 microsatellite primer pairs used in this study detected polymorphism with 2–6 alleles per locus. Similarity measures and cluster analysis were made using RAPD and simple sequence repeat (SSR) data, separately and together. The resulting dendrograms were compared with each other and with the available pedigree information as a control. The dendrogram derived from RAPD data showed some divergence from the pedigree information available for the lines. The dendrograms based on SSR data and SSR data combined with RAPD gave very good agreement with pedigree information. It can be concluded that the combined use of a limited number of RAPD and SSR markers is a useful and reliable means of evaluating genetic relationships of genotypes in the absence of pedigree data.     

具有中国大豆遗传背景的7S与11S多亚基缺失型大豆新品系的创制

利用我国高油大豆品种东农47与日本引进多亚基缺失型育种材料日B,采用回交、三交的育种方法,综合系谱选择,通过SDS-PAGE技术分析亚基组成,在BC₁、BC₃及三交种F₈群体内,选育到(α+11S groupⅡa)-缺失型、(α′+11S groupⅡa)-缺失型、[(α′+α)+11S groupⅡa、Ⅱb]-缺失型、[(α′+α)+11S groupⅡa]-缺失型、[(α′+α)+11S groupⅡb+X₁X₂]-缺失型、[(α′+α)+11S groupⅡb]-缺失型和(α′+11S groupⅠ、Ⅱa)-缺失型共7种具有中国大豆遗传背景的7S球蛋白α′、α亚基与11S球蛋白groupⅠ(A_1aB_1b,A₂B_1a,A_1bB₂)、groupⅡa(A₄A₅B₃)和groupⅡb(A₃B₄)不同亚基缺失组合新种质。测定优良品系的综合农艺性状及氨基酸组成、含量,结果表明,与对照相比,各种缺失突变体的各种氨基酸组分含量普遍提高,蛋白总量普遍高于轮回亲本,精氨酸含量特别是游离精氨酸的含量大幅提高。其中亚基组成为(α+11S groupⅡa)-缺失型品系G2-2-3的17种氨基酸含量、氨基酸总量、蛋氨酸含量均显著高于轮回亲本东农47,特别是游离精氨酸含量高出7.27mg/g。以上结果表明,7S与11S多亚基缺失型优良品系在有效去除致敏蛋白的同时,可以提高大豆蛋白氨基酸含量, 改善大豆蛋白氨基酸组分配比。各种致敏蛋白缺失型大豆优良新品系的获得,大大丰富了我国蛋白质组分改良育种的种质基础。     

Integrating marker assisted background analysis with foreground selection for identification of superior bacterial blight resistant recombinants in Basmati rice

Basmati rice is highly susceptible to bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae. Transfer of BB resistance genes from non‐Basmati sources to Basmati through cross‐hybridization requires strict monitoring for recovery of the desirable Basmati quality traits in the recombinants, which show complex inheritance pattern. We integrated background analysis using mapped microsatellite markers with foreground selection to identify superior lines that combine useful genes from a non‐Basmati BB resistance donor line IRBB55 with grain and cooking quality characteristics of the popular Basmati rice variety ‘Pusa Basmati 1’ (PB 1) employing backcross pedigree strategy. Foreground selection using linked markers ensured presence of two genes, xa13 and Xa21 for BB resistance from IRBB55, and the recurrent parent PB 1 allele for the waxy locus giving intermediate amylose content and maintainer allele at fertility restorer locus in the BC₁F₅ recombinants. Background analysis enabled selection of recombinants with recurrent parent genome to the extent of 86.3% along with the quality traits. The extent of introgression of non‐Basmati donor chromosome segments in the superior selections was estimated to be < 7.8 Mb and < 6.7 Mb in the xa13 and Xa21 linked genomic regions, respectively. Association mapping identified three quantitative trait loci, one each for 1000‐grain weight, fertile grains/panicle and cooked kernel length. The backcross‐pedigree breeding strategy facilitated recovery of additional desirable characteristics from the donor in some of the selections. The elite selection Pusa 1460‐01‐32‐6‐7‐67 with maximum genomic background and quality characteristics of the recurrent Basmati parent gave resistance reaction against BB, similar to that of the non‐Basmati resistant check variety and recorded an yield advantage of 11.9% over the best check in the multiplication agronomic trial in the Basmati growing region of India. This line, which has been released as a new variety in the name of ‘Improved Pusa Basmati 1’ for commercial cultivation in India, is an example of successful application of marker assisted selection to variety development.