苜蓿耐盐基因分子标记的筛选

以耐盐苜蓿×敏盐苜蓿组合的F2群体为试验材料,利用改良BSA法筛选与苜蓿耐盐基因紧密连锁的分子标记.在对26组520条RAPD随机引物筛选中,共有66条引物为DNA多态性引物,选出一个与苜蓿耐盐基因相连锁的分子遗传标记.通过F2代群体的遗传分析,观测到分子标记与耐盐性等位基因之间发生重组,但重组值较小,在A8×D2杂交组合中,重组率为2.27%;在A5×D1杂交组合中,重组率为4.03%.这些结果表明,这一显性标记与苜蓿的耐盐基因座位连锁程度较为紧密.用国外登记的耐盐苜蓿种质及相对敏盐种质单株对获得的耐盐标记进行验证,85%耐盐种质材料AZ-90NDC-ST的单株DNA都扩增出1个约1 400bp的DNA片段;75%敏盐种质材料AZ-88NDC的单株DNA未能扩增出此片段.     

苹果砧木SRAP-PCR反应体系的优化及耐盐性标记的筛选

本研究以苹果砧木SH38为材料,利用正交试验设计对影响SRAP-PCR反应的4因素(Taq DNA聚合酶、模板DNA,d NTPs,引物)4水平进行优化,并构建苹果砧木SH38的SRAP-PCR的最佳反应体系。结果显示:15μL的总反应体系中,d NTPs浓度为0.3 mmol/L,Taq DNA聚合酶用量为0.75 U,上、下游引物为50 ng,DNA模板用量为60 ng。运用该反应体系并结合BSA法,从128对引物组合中筛选出4对引物,该4对引物扩增出的特异性片段与苹果砧木的耐盐基因存在连锁。研究结果可为苹果砧木利用SRAP分子标记进行耐盐性的分子标记辅助育种提供基础。     

苜蓿耐盐基因分子标记的筛选及鉴定

以耐盐苜蓿×敏盐苜蓿组合的F2群体为试验材料,利用改良BSA法筛选与苜蓿耐盐基因紧密连锁的分子标记。在对26组520条RAPD随机引物筛选中,共有66条引物为DNA多态性引物,选出一个与苜蓿耐盐基因相连锁的分子遗传标记。通过F2代群体的遗传分析,观测到分子标记与耐盐性等位基因之间发生重组,但重组值较小,在A8×D2杂交组合     

甜椒胞质雄性不育恢复系SRAP及SCAR标记

以甜椒胞质雄性不育恢复系5-2R1和相应的保持系5-2为试材,利用SRAP分子标记技术筛选与甜椒恢复基因相关的分子标记。128对SRAP引物共扩增获得了3796条从100bp至800bp大小不同的条带,平均每对引物组合可扩增出30条清晰的条带,检测到4个多态性位点,其中恢复系材料仅有1个。对该特异片段进行回收、克隆和测序,结果表明该片段全长为259bp。经数据库比对分析表明,该片段与线粒体中的NADH脱氢酶第5亚基(GenBank:EF151914.1)具有81%的同源性;同时,该片段与辣椒BAC克隆PEPBAC 158K24(GenBank:FJ597540.1)的部分序列高度同源。根据序列特点设计特异SCAR引物,对恢复系和保持系进行扩增验证,仅在恢复系中扩增出目的条带,表明已成功地将此SRAP标记转化为简单、稳定的SCAR标记。我们推测本研究获得的SCAR标记可能与胞质雄性不育恢复性状连锁。     

瓠瓜耐湿涝相关根系性状遗传规律及分子标记筛选

以耐湿涝瓠瓜材料JZS和不耐湿涝材料T2002及其杂交F2群体为材料,选择湿涝条件下不定根数目作为耐湿涝鉴定指标,分析瓠瓜耐湿涝特性遗传规律,并采用集团混合分离分析法筛选相关分子标记。结果表明,瓠瓜亲本JZS的耐湿涝特性由1对显性单基因控制,F1的不定根数目比耐湿涝亲本JZS增多,表现出超亲优势。在600个RAPD和100对瓠瓜SSR引物中,共有18个引物(14个RAPD和4对SSR)在双亲中显示多态性。最终筛选到瓠瓜SSR引物S87/88,在双亲及基因池间稳定扩增出片段大小为230 bp的差异条带。通过F2153个单株验证,确定表型调查耐湿涝结果与分子标记S87/88鉴定结果相关性极显著(相关系数0.78),利用Mapmaker 3.0软件对分子标记与表型结果进行连锁分析,S87/88与瓠瓜耐湿涝相关的不定根数目基因的连锁距离为8.8 cM,为进一步利用分子辅助育种加速耐湿涝瓠瓜新品种的选育提供有效标记。     

甘薯SRAP连锁图构建淀粉含量QTL检测

SRAP标记（sequence-related amplified polymorphism,序列相关扩增多态性）是由Li和Quiros发展了一种新的分子标记技术.SRAP标记具有简便、稳定、中等产率、在基因组中分布均匀的特点,已被广泛利用.本实验利用甘薯高淀粉品种绵粉一号与甘薯低淀粉品种红旗四号杂交F1代分离群体,根据淀粉含量,选择F1代分离群体中高、低淀粉含量极端类型材料各23个构成选择性基因型子群体.构建的连锁图包括21个SRAP标记和9个连锁群（母本4个连锁群,父本5个连锁群）.复合作图检测到1个淀粉含量QTL,红旗4号的加性效应增加淀粉6.37%,解释表型变异的20.1%.     

水稻地方品种苗期耐盐QTL的定位

水稻耐盐遗传位点的发掘可为耐盐遗传机制的研究提供理论基础,为耐盐品种培育提供基因资源。以云南地方籼稻品种扎西玛与江苏著名优质粳稻品种南粳46为亲本构建的水稻重组自交系群体为研究对象,鉴定了各株系的苗期耐盐性,结合该群体的分子连锁图谱对控制水稻苗期耐盐性QTL进行分析,共检测到4个QTLs:qSST-1、qSST-3、qSST-5和qSST-11,分别位于第1,3,5,11号染色体上。4个QTLs的增效等位基因均来自于亲本南粳46。经比较发现有3个QTLs与已克隆水稻耐盐基因不在同一染色体区间,说明为新的耐盐基因候选位点。结果对进一步发掘和利用新的水稻耐盐QTL具有重要意义。     

与甜瓜嫁接亲和性连锁分子标记鉴定

使用与南瓜类型砧木(“壮士”)嫁接亲和性差异明显的两个甜瓜自交系(85-1和B717)为亲本,配制F2∶3家系群体;利用BSA法构建亲和性优差池,结合PCR技术筛选与嫁接亲和性有关的分子标记.结果表明:使用葫芦科遗传连锁图谱上发表的750对SSR、SRAP和AFLP等分子标记引物扩增,最终筛选出在两个甜瓜材料之间具有多态性的382对引物,其中9对引物分别在嫁接亲和性优差DNA池中扩增出差异性片段.将这些标记用于F2群体的扩增,计算标记与嫁接亲和性遗传连锁距离,两对SSR引物CMN-0616和M103与嫁接亲和性的遗传距离分别为17.6 cm和9.3 cm.这2对引物有望成为鉴定甜瓜嫁接亲和性的辅助引物,从而将嫁接亲和性差异材料从分子角度区别开来,为甜瓜嫁接接穗育种等研究提供参考.     

杏鲍菇遗传多样性的SRAP和ITS分析

研究来自中国不同地区和日本的27株杏鲍菇栽培菌株的遗传多样性,以期为杏鲍菇的鉴定和选育提供分子依据。利用SRAP和ITS标记联合使用的方法,通过聚类分析对供试杏鲍菇菌株进行研究。筛选出9对SRAP引物共扩增出151条条带,其中具有多态性条带130条,平均多态性比例为83.3%,多态性信息含量(PIC)变幅在0.27~0.42,平均为0.36。通过ITS序列对供试菌株进行亲缘关系分析,与SRAP分析的结果一致,均表明地域来源相近的部分菌株聚在一起,亲缘关系较近,而地域相隔较远的部分菌株也聚为一类,其亲缘关系也很近。2种标记均显示供试杏鲍菇栽培菌株的遗传多样性较为丰富,其遗传相似性与地理分布存在一定的联系。SRAP和ITS联合使用能够得到更好的效果,从而为杏鲍菇遗传多样性研究提供更为有力的参考。     

一个与亚洲百合无花粉性状紧密连锁的SRAP标记

为了探讨亚洲百合无花粉性状的分子遗传机制,筛选与该性状紧密连锁的分子标记,实现分子辅助选择。本研究以无花粉与有花粉亚洲百合杂交创制的F1单株为试验材料,采用BSA法,对亚洲百合无花粉性状进行SRAP标记分析。在无花粉基因池和有花粉基因池之间共筛选了108对SRAP引物组合,获得了1对在两基因池间扩增存在显著差异的引物组合EM2/ME3,并成功对两基因池间扩增出的差异片段进行回收、克隆和测序,得到了该片段375 bp的碱基序列,并将该特异标记命名为E2M3-375。     

Biological Activity and Quantification of Suspected Allelochemicals from Alfalfa Plant Parts

Autotoxicity restricts reseeding of alfalfa (Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments. A series of aqueous extracts from leaves, stems, roots and seeds of alfalfa ‘Vernal’ were bioassayed against alfalfa seedlings of the same cultivar to determine their autotoxicity. The highest inhibition was found in the extracts from the leaves. Extracts at 40 g dry tissue l^?1 from alfalfa leaves were 15.4, 17.5 and 28.7 times more toxic to alfalfa root growth than were those from roots, stems and seeds, respectively. A high‐performance liquid chromatography (HPLC) analysis with nine standard compounds showed that the concentrations and compositions of allelopathic compounds depended on the plant parts. In leaf extracts that showed the most inhibitory effect on root growth, the highest amounts of allelochemicals were detected. Among nine phenolic compounds assayed for their phytotoxicity on root growth of alfalfa, coumarin, trans‐cinnamic acid and o‐coumaric acid at 10^?3 m were most inhibitory. The type and amount of causative allelochemicals found in alfalfa plant parts were highly correlated with the results of the bioassay, indicating that the autotoxic effects of alfalfa plant parts significantly differed.     

Changes in Seed Yield and Quality with Maturity in Onion (Allium cepa L., cv. 'Early Cream Gold')

Development of onion (Allium cepa L., cv. ‘Early Cream Gold’) seed under cool climate conditions in Tasmania, Australia occurred over a longer duration than previously reported, but similar patterns of change in yield components were recorded. In contrast to previous studies, umbel moisture content declined from 85 to 67 % over 57 days while seed moisture content decreased from 85 to 31 %. Seed yield continued to increase over the duration of crop development, with increasing seed weight compensating for seed loss resulting from capsule dehiscence in the later stages of maturation. Germination percentage was high and did not vary significantly from 53 to 77 days after full bloom (DAF), but mean germination time declined and uniformity of germination increased significantly over the same time period. The percentage abnormal seedlings declined with later harvest date, resulting in highest seed quality at 77 DAF. The results of this study suggest that the decision to harvest cool climate onion seed crops before capsule dehiscence will result in a loss of potential seed yield and quality.     

Location of a high-lysine gene and the DDT-resistance gene on barley chromosome 7

Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans.     

A note on the origin of Kalanchoë x vadensis Boom & Zeilinga (Crassulaceae)

Summary K x vadensis is a hybrid of K. blossfeldiana and K. marmorata obtained after doubling the number of chromosomes.     

Simultaneous Determination of Four Phenolic Acids in Radix Salviae Miltiorrhizae Formula Granules by QAMS

[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol...     

Avoidance of leaf rust fungi in wild relatives of cultivated cereals

Summary Avoidance of rust fungi that was based on poor appressorium induction was previously found in Hordeum chilense. In the present study 95 accessions of Triticeae were screened for avoidance of Puccinia hordei. The percentage of appressorium formation per germinated spore ranged from 6 to 90%. On none of the 41 accessions of Aegilops, Agropyron, Elymus, Secale, Thinopyrum or Triticum studied was the rate of appressorium formation lower than 25%. Lower rates of appressorium formation were, however, found on accessions of wild barley species Hordeum brachyantherum, H. marinum, H. parodii and H. secalinum. Its implications in cereal breeding are discussed.     

Variation in forage grasses as hosts of the root-knot nematode Meloidogyne naasi (Franklin) and selection for resistance in ryegrass

Summary Reactions of 13 grasses to Meloidogyne naasi varied with species; ryegrasses, fescues and their hybrids were generally susceptible and cocksfoot and timothy resistant. Marked variation in host resistance levels occurred between genotypes within cultivars.Selection of single plants, followed by tests on replicate tillers, identified resistant and susceptible genotypes in both Italian and perennial ryegrass cultivars. Resistant plants had few nematode-induced galls and fewer females and eggs than susceptibles. There was more or less continuous variation, with many genotypes intermediate between extremes of resistance and susceptibility. Selected resistant and susceptible genotypes are of use in assessing variation in nematode populations and as controls for breeding and selection programmes.     

Current and future strategies in breeding lentil for resistance to biotic and abiotic stresses

Lentil production is limited by lack of moisture and unfavorable temperatures throughout its distribution. Waterlogging and salinity are only locally important. Progress has been made in breeding for tolerance to drought through selection for an appropriate phenology and increased water use efficiency and in breeding for winter hardiness through selection for cold tolerance.The diseases rust, vascular wilt, and Ascochyta blight, caused by Uromyces viciae-fabae, Fusarium oxysporum f. sp. lentis, and Ascochyta fabae f. sp. lentis, respectively, are the key fungal pathogens of lentil. Cultivars with resistance to rust and Ascochyta blight have been released in several countries and resistant sources to vascular wilt are being exploited. Sources of resistance to several other fungal and viral diseases of regional importance are known. In contrast, although the pea leaf weevil (Sitona spp.) and the parasitic weed broomrape (Orobanche spp.), and to a lesser extent the cyst nematode (Heterodera ciceri), are significant yield reducers of lentil, no sources of resistance to these biotic stresses have been found. Directions for future research in lentil on both biotic and abiotic stresses are discussed.     

Breeding forage grasses for organic conditions

Herbage yield trials are necessary to test if candidate varieties are well suited for organic conditions. In order to elucidate a particular suitability for organic conditions, we run such trials in parallel under both organic and conventional conditions. Here, we report on second and third year results of the sowings 2004 and 2005. These trials are part of a targeted breeding programme for organic conditions. Irregular colonisation of pure grass plots with adventitious plants, in particular white clover, was recognized as a major constraint to the quality of the results obtained under organic conditions. The coefficients of variance in dry matter yield almost doubled when compared to conventional trials, and significant differences in annual yield among varieties were rare. However, when herbage yield was corrected for the estimated fraction of adventitious plant colonization at each cut, the number of instances with statistically significant differences for annual yield among varieties increased more than fourfold. Dry matter yields under organic and conventional conditions were significantly and positively correlated. However, analysis of variance showed a significant culture type by variety interaction for annual dry matter yield in 12 out of 24 trials. When diploid and tetraploid varieties of the same Lolium species were tested in one common trial, only tetraploid varieties were identified as being particularly well suited for organic conditions, while in these trials, the great majority of apparently poorly suited varieties were diploid. This suggests that breeding tetraploid Lolium varieties is promising when aiming at a favourable response to organic agriculture.     

The performance of double or mixed pollinations with compatible and self-incompatible or incongruous pollen of pear and apple

Summary Six pear and five apple trials were carried out to ascertain the outcome of combinations of compatible pollen (C) with self (S) or incongruous pollen (I) as to the pollination index (PI=seeds/pollinated flower). The PI of the mixture C+I (1:5) was consistently depressed as compared to that of the control C. The results of the double pollinations S/C and I/C were affected by the temperature at pollination; their PI's at <15°C were twice as high as those at >15°C, being well above and below the PI of C in the former and latter case respectively. The opposite was true for the C/S combination, the PI of which increased with the pollination temperature; the PI of C/I did not differ much from the PI of C, irrespective of temperature. The conclusion was reached that the interaction previously and presently found between compatible and self-incompatible pollen also exists to a fair extent between compatible and incongruous pollen. However, in pear neither the mentor nor the pioneer pollen technique proved to aid its hybridization with apple, the formation of self seed was not observed either. In apple the production of apple × pear hybrids was likewise doubtful, but the double pollinations S/C and C/S formed 4–10% self seed.