小麦株高近等基因系的RAPD标记研究

采用ＲＡＰＤ技术，以４个小麦株市基因近等基因系（分别含有rht、Ｒht1、Rht2、Rht3)为材料，对２９６个单一随机引物（１０个核苷酸）进行了筛选。发现２５个引我的扩增产物的近等基因系间表现出特异性，在６次重复试验中，有１８个引物的特异护增片段不能重复，６个引物可以重复２－３资，唯有ＯＰＡＭ０１在全部试验中均能稳定重复，其特异扩增版段ＯＰＡＭ０１１８６０可以作为rht基因的ＲＡＰＤ标记。     

不同Rht基因对小麦子粒品质的影响

多年结果证明Rht1、Rht2、rht3个近等基因系之间的子粒蛋白质含量差异不显著，而Rht3基因系的蛋白质含量显著高于前三者。Rht3基因系的子粒赖氨酸含量显著高于Rht2和rht系统，Rht10显著高于Rht1系，其他基因系之间无显著差异。Rh10基因系的面粉的沉降值显著高于Rht1,Rht8和Rht12,Rht3显著高于Rht1,Rht2和rht。Rht10和Rht3基因系面粉的湿面筋显著高于其他基因系。此外，同一基因系在不同的年限之间的品质性状表现有差异。     

春小麦和硬粒小麦主效矮秆基因近等基因系的全球适应性

小麦和硬粒小麦中Rht—B1和Rht—D1主效矮秆基因的效应随环境而变化。我们在全球81个试验中种植了国际适应性试验（IAT）中的6个春小麦矮秆近等基因系。在产量大于3Mg／ha的56个试验中，有54％的试验的半矮秆品系平均产量显著高于高秆品系；在产量低于3Mg／ha的27个试验中，仅有在24％的试验中为半矮秆品系的产量占优势。     

四倍体小麦矮秆基因的赤霉素敏感性及对农艺性状的影响

为明确四倍体小麦矮秆基因Rht14、Rht16和Rht18的赤霉素敏感性及其对小麦农艺性状的影响,促进这些矮秆基因的合理利用。选用分别含有Rht14、Rht16和Rht18的四倍体小麦近等基因系ANW16D(Rht14)、ANW16F(Rht16)、ANW16G(Rht18)及其高秆轮回亲本LD222以及六倍体小麦中国春(Chinese spring),测量其不同浓度GA3处理下小麦的株高,计算赤霉素敏感系数(GRI)并推断3种矮秆小麦的赤霉素反应类型。在成熟期对LD222近等基因系小麦的农艺性状如株高、穗长、主穗穗下第一茎节(P-1)节间长、节间表皮细胞、种子表皮细胞及种子体积等进行测量,分析Rht14、Rht16、Rht18这3个矮秆基因对小麦这些农艺性状的效应。结果表明,ANW16D、ANW16F和ANW16G这3个矮秆小麦株高恢复到正常LD222株高的最适GA3浓度为10-4mol/L;3个矮秆品种均为赤霉素敏感型且敏感性大小为中国春ANW16GANW16FANW16DLD222;Rht14、Rht16、Rht18均是通过降低主穗穗下第一茎节(P-1)节间长度来使小麦株高降低,降低效应为Rht18Rht16Rht14;它们降低小麦株高的根本原因均是缩短了小麦节间表皮细胞长度且缩短效应与降低株高效应一致;3个矮秆基因均在降低小麦株高的同时不影响种子的体积。     

小麦显性矮秆基因Rht3理想株高突变体的基因型检测

本研究采用4BS染色体携带Rht3基因的小麦显性矮源"矮苏3"(55～60 cm),经辐射与化学诱变,获得了一系列株高在70～85 cm、具小麦育种理想株高的突变体.采用形态标记、生化标记及分子标记对上述理想株高突变体进行了基因型检测.经成熟种子萌发试验的生化标记检测表明,理想株高突变体仍具有显性矮秆基因Rht3成熟种子α-淀粉酶活性低而抗穗萌的特性.经采用位于4BS染色体上的"易组太谷核不育基因MS2 (4BS)"作为形态标记基因来定位理想株高突变体携带的半显性矮秆基因,证实了理想株高突变体携带的半显性矮秆基因与MS2(4BS)连锁、因而与Rht3基因同位于普通小麦4B染色体上.基于通常认为Rht3与隐性矮秆基因Rht1同为4BS染色体上的复等位基因,经采用Ellis等开发的"perfect marker"SSR特异引物的分子标记检测,在矮苏3及其理想株高突变体上同时扩增出了与Rht-B1b相同的237 bp的特征带.以上3种类型的基因标记检测的结果,均有利于说明矮苏3的理想株高突变体携带Rht3突变衍生的复等位基因,因其具理想株高而又抗穗萌,可望作为半显性创新矮源用于高度集约化的小麦"分子设计育种",以克服小麦育种目前局限于使用隐性矮源的局面,实现自"绿色革命"以来小麦育种矮源的升级换代.     

甘蔗线虫病抗性基因的PCR检测研究

以38份未经线虫病常规鉴定的甘蔗种质资源为供试材料,根据番茄抗根结线虫病基因和甜菜抗胞囊线虫病基因的保守序列区域,分别设计了2条上游引物、2条下游引物,对设计的引物进行组合后,应用PCR特异扩增从中分别筛选出各1对特异引物。用抗根结线虫基因设计的特异引物进行扩增最终获得了1条大约460 bp大小的特异片段;用抗胞囊线虫基因设计的特异引物进行扩增最终获得了1条大约690 bp大小的特异片段,进一步进行了PCR-Southern杂交,确认了该2条特异引物的真实性。     

山东小麦品种中矮秆基因Rht-B1b、Rht-D1b分布的分子鉴定

利用小麦矮秆基因Rht—B1b、Rht—D1b的4对特异分子标记,NH—BF和MR1、NH—BF和WR1．2、DF和MR2、DF和WR2,对山东小麦品种中矮秆基因Rht—B1b及Rht—D1b的分布情况进行了分子标记鉴定,结果表明在鉴定的150个品种中有20．67％含有Rht—B1b基因,有54．00％含有Rht—D1b基因,二者合计为74．67％;同时含有Rht—B1b和Rht—D1b2个矮秆基因的仅占3．33％。表明山东小麦矮化育种中Rht—D1b基因的利用远高于Rht—B1b。     

鉴别牛早期胚胎性别的多重PCR技术研究

根据牛Y染色体特异重复序列设计合成5对牛Y染色体特异引物,依据牛微卫星基因设计合成3对牛染色体内标引物.每对引物分别扩增牛基因组DNA,筛选出2对牛Y染色体特异引物和1对牛DNA特异内标引物.通过对多重PCR扩增条件进行优化组合,建立1个可用于牛早期胚胎性别鉴别的多重PCR方法及引物组合:BY1/A1.     

大白菜细胞质雄性不育的分子鉴定及序列分析

为了获得3种大白菜细胞质雄性不育系Ogu CMS,Pol CMS,CMS96和保持系间的多态性以及定位大白菜CMS96不育系所属的不育类型,利用设计的atp6,orf222单一和混合引物PCR扩增3组11份同核异质大白菜细胞质雄性不育系和保持系mtDNA。结果表明,atp6引物在大白菜Ogu CMS不育系扩增的200 bp片段为其特异带;orf222引物仅在大白菜Pol CMS和CMS96不育系有扩增产物,但二者有3点完全不同:大白菜Pol CMS不育系扩增产物为675bp,CMS96不育系扩增产物为669 bp,二者相差6个核苷酸,后者定名为大白菜CMS96-orf222。大白菜CMS96-orf222与甘蓝型油菜Nap CMS的nad5c基因和Nap-orf222基因同源性均为99%,E值为0.0;大白菜Pol CMS的675 bp序列具有ORF224开放阅读框,没有保守结构域,而大白菜CMS96的669 bp序列具有ORF222开放阅读框和保守结构域YMF19。另外,atp6和orf222混合引物多重PCR扩增产物存在明显多态性:800 bp为大白菜保持系的差异带型;2 300 bp和1 500 bp为大白菜Pol CMS不育系特异带型;200 bp为大白菜Ogu CMS不育系特异带型;690 bp为大白菜CMS96不育系特异带型。该方法仅用一次PCR反应快速地将3种大白菜细胞质雄性不育系和保持系一次性全部区分开,为大白菜分子育种和常规育种更好地相结合提供了简单、快速、准确和重复性好的方法和手段。     

N基因标记基因PCR检测方法的建立及其在遗传育种中的应用

烟草的N基因是一个较为有效的抗烟草花叶病毒(tobacco mosaic virus,TMV)基因,在抗病育种中发挥着重要的作用。为建立其标记基因PCR检测体系,评估已知目的基因cDNA序列作为分子标记的目的基因辅助育种的可行性,本文依据N基因cDNA序列设计4对扩增范围在150～950bp的不同片段长度引物,并对含有N基因的coker176基因组DNA进行PCR扩增。扩增结果表明,有3对可扩增出特异产物。测序结果表明,有2对引物的扩增产物含有内含子。根据所获得的DNA序列设计出2对用于分子标记辅助育种的引物,并分别对C151、NC89、coker176等11个品种的基因组DNA进行扩增,结果表明在coker176、龙江912、吉烟9号和龙江911品种中有扩增产物,除龙江911外,这一结果与已知的N基因在11个品种的分布相一致,成功建立了N基因PCR检测体系,并证明以cDNA序列设计引物扩增DNA序列的目的基因辅助育种技术完全可行。并对实验中出现的问题进行了研究。     

Effects of dwarfing genes on yield and yield components under irrigated and rainfed conditions in wheat (Triticum aestivum L.)

Summary Near-isogenic tall (no dwarfing gene), semidwarf (Rht1 or Rht2) and dwarf (Rht1 + Rht2 or Rht3) spring wheat lines were evaluated for yield and yield components under irrigated and rainfed conditions. Under irrigated conditions, the dwarf and the semidwarf lines exhibited a significant yield advantage over the tall lines. Under rainfed conditions, the semidwarf lines outyielded the tall as well as the dwarf lines. Percent yield reduction in response to drought stress was highest with the dwarfs and lowest with the tall lines. Dry matter production of the tall lines and that of the semidwarf lines did not differ significantly and both produced significantly more dry matter than the dwarf lines under irrigated as well as rainfed conditions. Plant height and kernel weight decreased with increasing degree of dwarfness while number of kernels per spikelet, harvest index and days to heading increased under both moisture regimes. The dwarfing genes did not have any significant influence on number of tillers/m² and spikelets per spike in either moisture regime.     

人工合成小麦与普通小麦杂交后代衍生群体的Rht8基因分析

【目的】四倍体小麦与节节麦杂交培育的人工合成小麦已广泛应用于国内外小麦品种改良。通过研究人工合成小麦与普通小麦杂交后代的Rht8基因型,有助于提高分子标记育种效率,也有助于Rht8 基因型的多态性研究,并为人工合成小麦在中国小麦品种改良和分子标记育种中的应用提供依据和方法;【方法】以引自CIMMYT的人工合成小麦分别与中国四川成都平原主栽普通小麦品种杂交、回交的BC2F2:6后代群体中选育的113份优良高代系和川麦38、川麦42、川麦43和川麦47育成品种为材料,采用特异引物的PCR 扩增和改进的聚丙烯酰胺凝胶电泳对其Rht8基因型进行了研究;【结果】在以syn768、Syn769、Syn780和Syn786人工合成小麦为亲本的117份后代衍生群体检测材料中,Rht8基因型频率为77.78%。从每一个人工合成小麦形成的小的后代衍生群体看,Rht8基因型频率各不相同。以syn768为亲本的后代衍生群体,Rht8基因型频率最高,为96.70%;在以syn769为亲本而育成的优良高代系和川麦38、川麦42与川麦43育成品种中,Rht8基因型频率最低,为71.64%;以Syn780为亲本的后代衍生群体中,Rht8基因型频率为73.68%,分离比率约为3:1;以Syn786为亲本育成的材料只有川麦47,该品种不含有Rht8该基因;【结论】不论父本或母本的Rht8的基因型状况如何,它们所产生的杂交后代材料Rht8基因的遗传是随机的。     

Classification of GA response,Rht genes and culm length in Japanese varieties and landraces of wheat

Summary The GA response, Rht genes and culm length of 133 Norin varieties, 6 breeding lines and 16 landraces of Japanese wheat were investigated. Out of 133 Norin varieties tested, 103 were GA-insensitive and 30 GA-responsive. The 6 breeding lines were all GA-insensitive. Out of 16 landraces tested, 10 were GA-insensitive and 6 GA-responsive. Among the 10 GA-insensitive landraces, only Daruma had a Rht1 genotype. The other 9 had a Rht2 genotype. None of the landraces tested carried both Rht1 and Rht2 or Rht3. Out of the 103 GA-insensitive Norin varieties, 22 carried only Rht1, another 79 carried only Rht2, and only Norin 10 and Kokeshikomugi carried both Rht1 and Rht2. No tested variety carried Rht3. Some Norin varieties carrying Rht2 showed tall culms comparable to that of the rht tester line Chinese Spring. These results suggest that these varieties had a nullifier or modifier gene(s) or height promoting genes in the background controlling the height-reducing effect of Rht2. Conversely, six GA-responsive Norin varieties were as short as Akakomugi which carries the GA-responsive Rht genes, Rht8 and Rht9. The also seemed to carry a GA-responsive Rht gene or genes, and moreover all but one may carry gene(s) other than the Akakomugi genes. The origin of Rht1 and Rht2 of Norin 10 was speculated on the GA-response and Rht genotypes of its related varieties and landraces.     

Pleiotropic Effects of Genes for Reduced Height (Rht) and Day-Length Insensitivity (Ppd) on Yield and its Components for Wheat Grown in Middle Europe

Under field conditions in Germany over three growing seasons the pleiotropic effects on yield and its components of four sets of near isogenic lines carrying the GA insensitive dwarfing alleles Rht1, Rht2, Rht3, Rht1+2, Rht2+3 or rht (tall) in four different genetical backgrounds were examined together with 24 single chromosome recombinant lines segregating for the GA sensitive dwarfing gene Rht8 and the gene for day-length insensitivity Ppd1 in a ‘Cappelle-Desprez’ background. For the GA insensitive semi-dwarfs it was shown that in all three years a higher number of grains per ear was accompanied by a lower grain weight. Depending on the climatic conditions in a particular year, the increase in grain number was sufficient to compensate for the reduction in grain size and resulted in higher yields. For the Ppd1 allele yield advantages were found for wheats grown under environmental conditions of middle Europe.     

The effect of major dwarfing genes on yield potential in spring wheats

Summary A composite convergent cross of 16 spring wheat parents produced a set of unselected progeny lines among which the major dwarfing genes, Rht1, Rht2 and Rht3, were distributed against a common random genetic background. Random subsets of these lines were grown under irrigation and optimal conditions in 4 experiments with replicated bordered plots in southern New South Wales in order to measure the dwarfing gene effect on yield potential. The dwarfing gene composition of each line was determined by test crossing and seedling responsiveness to gibberellic acid.Lodging was negligible in the two experiments in 1982. While present in the two in 1983, it was not strongly associated with yield. Grain yield levels were appropriately high (mean 5.9 t/ha). In all but 1 experiment the Rht1+Rht2 dwarf genotypes gave highest yields while the Rht3 group yielded on average 3% lower, Rht2 9% lower, Rht1 11% lower, and the non-dwarf or tall group yielded 24% lower. These yield differences were positively associated with harvest index, kernels per m² and kernels per spike, but negatively associated with mature plant height. Even within major dwarfing gene classes, grain yield was significantly and negatively associated with height.     

Rht8、Rht10和Rht12矮秆基因对产量构成因子的影响

利用携带不同矮秆基因的近等基因系,通过两年、两地的试验研究证明,Rht8半矮秆基因虽然其总小穗数显著低于其他系,但其籽粒产量、小花结实率较Rht10和Rht12显著高.Rht10的降秆作用最强,千粒重显著高于Rht8,但其分蘖成穗率、结实率及小区籽粒产量显著低于其他,表现对环境条件特敏感.Rht12矮秆基因的降秆程度显著高于Rht8,但由于生物产量太低、成熟太晚,造成籽粒产量显著降低,在小麦育种中单独利用价值较低.     

矮杆基因对春小麦植株生长发育的影响及对子粒产量的间接作用

利用以春小麦品种ＡｐｒｉｌＢｅａｒｄｅｄ为背景的含有不同矮秆基因Ｒｈｔ１、Ｒｈｔ２、Ｒｈｔ３、Ｒｈｔ１＋Ｒｈｔ２和Ｒｈｔ２＋Ｒｈｔ３的５个近等基因系,研究了不同矮秆基因对小麦生长发育的作用。结果表明：Ｒｈｔ１半矮秆基因显著缩短了小麦植株生长发育进程,促进了地上部干物质积累,加大了旗叶面积和倒二叶面积,提高了单株成穗率,减少了无效分蘖;Ｒｈｔ２半矮秆基因显著增大了倒二叶面积,提高了单株成穗率和穗长,减少了无效分蘖;Ｒｈｔ３矮秆基因显著加大了旗叶面积,但对地上部干物质的积累、单株成穗和穗长均有显著的负向作用;Ｒｈｔ１＋Ｒｈｔ２基因结合没有突出的优势存在：Ｒｈｔ２和Ｒｈｔ３基因结合对有利于提高小麦产量性状的作用均为负向最大;上述３种矮秆基因及其不同的结合形式均有显著的使茎秆矮化的作用。因此认为Ｒｈｔ１半矮秆基因在小麦育种中利用价值较大,Ｒｈｔ３矮秆基因利用价值则较小。     

不同矮秆基因对冬小麦农艺性状的影响

利用以冬麦品种ＭａｒｉｓＨｕｎｔｓｍａｎ为背景的含有不同矮杆基因Ｒｈｔ１,Ｒｈｔ２和Ｒｈｔ３及其不同结合形式的６个近等基因系,研究了不同矮秆基因对小麦农艺性状的作用,结果表明,Ｒｈｔ１半矮秆基因显著提高了单株穗数,粒数和粒重,地下部生物产量,经济系数和倒二叶面积,Ｒｈｔ２半矮秆基因显著提高了单株（或单穗）粒数和粒重,经济系数和倒二叶面积,显著降低了千粒重,Ｒｈｔ３矮秆基因对单株粒数,地上部生物产量     

Genotype analysis of the wheat semidwarf Rht‐B1b and Rht‐D1b ancestral lineage

In wheat, semidwarfism resulting from reduced height (Rht)‐B1b and Rht‐D1b was integral to the ‘green revolution’. The principal donors of these alleles are ‘Norin 10’, ‘Seu Seun 27’ and ‘Suwon 92’ that, according to historical records, inherited semidwarfism from the Japanese landrace ‘Daruma’. The objective of this study was to examine the origins of Rht‐B1b and Rht‐D1b by growing multiple seed bank sources of cultivars comprising the historical pedigrees of the principal donor lines and scoring Rht‐1 genotype and plant height. This revealed that ‘Norin 10’ and ‘Suwon 92’ sources contained Rht‐B1b and Rht‐D1b, but the ‘Seu Seun 27’ source did not contain a semidwarf allele. Neither Rht‐B1b nor Rht‐D1b could be definitively traced back to ‘Daruma’, and both ‘Daruma’ sources contained only Rht‐B1b. However, ‘Daruma’ remains the most likely donor of Rht‐B1b and Rht‐D1b. We suggest that the disparity between historical pedigrees and Rht‐1 genotypes occurs because the genetic make‐up of seed bank sources differs from that of the cultivars actually used in the pedigrees. Some evidence also suggests that an alternative Rht‐D1b donor may exist.