基于松树EST序列的马尾松SSR引物开发

本文用生物信息学方法对松树359 521条EST进行处理,得到无冗余EST序列56 776条,其中有2 315个SSR分布于2 057条EST中,出现频率是4.08%,平均距离是22.06 kb.检测到二、三碱基重复SSR总数为1 631个,四碱基至六碱基重复SSR共117个.在所得二碱基以上重复SSR中,多态潜能高的有488个,占27.92%;多态潜能次高的有1 260个,占72.08%.选择整体长度不小于24个碱基的SSR位点用于开发马尾松SSR引物,设计出135对备选引物,最终有51对引物在马尾松中得到扩增,有效扩增率为37.78%;其中39对有多态性,多态率为28.89%.在51对可扩增引物所对应的SSR类型中,P型占74.51%,I型占3.92%,C型占21.57%.     

鹅掌楸EST-SSR引物开发及通用性分析

通过对6520条鹅掌楸EST序列进行检索,在364条ESTs中发现394个SSRs,鹅掌楸EST-SSRs平均密度为每8.5kb含有1个SSR;在检索出的SSRs中,二核苷酸重复单元的SSRs类型最多,占总数的61.9%。利用SSR-ESTs序列共设计176对EST-SSR引物,其中132对在鹅掌楸上有扩增产物,66对扩增出多态,多态性引物占所设计引物的36.9%。这批EST-SSR引物在物种之间具有较高的通用性。研究表明在鹅掌楸中表现多态的66对EST-SSR引物,85%在中国马褂木中有扩增,54%在白玉兰中有扩增。     

大白菜抗芜菁花叶病毒(TuMV)EST-SSR信息分析与引物筛选

为了研究大白菜抗芜菁花叶病毒(TuMV),对大白菜及芸薹属中与抗性有关的EST序列进行了搜索和SSR筛选,并根据SSR两端的序列设计引物,以1对大白菜TuMV抗、感材料的基因组DNA为模板,对上述引物进行了筛选。结果表明,在132条大白菜EST和895条芸薹属抗性EST序列中,共筛选出166个符合条件的SSR(16.2%)。其中包含6种核苷酸重复基元,占主导地位的分别是二核苷酸重复(33.7%)和三核苷酸重复(50.0%),其余重复类型所占比例较小。根据上述SSR两翼的序列,共设计引物196对。引物筛选结果表明,能够进行成功扩增的有156对,占79.6%,其中35对引物在2份材料中具有多态性。因此,利用芸薹属中抗性有关的EST序列设计SSR引物,可用于大白菜TuMV抗性有关基因的标记。     

棉花非冗余性EST-SSR新标记的开发及其评价

利用Clustal X等软件对公共数据库现有的393 753条棉花EST序列分析,得到349 815条非冗余EST序列,借助自主开发的SSRmine软件共发掘SSR位点11 372个,分布于10 507条EST中,EST-SSR的频率是3%,平均相隔21 kb出现一个SSR。在2~6 bp的重复基元中,三核苷酸和六核苷酸分别占34.1%、40.6%,二、三、四、五和六核苷酸基序分别以AG/CT、AAG/CTT、AAAT/ATTT、AAAAG/CTTTT和AAAAAG/CTTTTT的类型最多。利用去冗余的且在亚洲棉、陆地棉、海岛棉中没有被开发过的410条EST序列设计开发了200对非冗余性SSR引物,利用自主开发的SSRD软件通过SSR引物序列下载、预处理、Blastn、提取相似性分值≥81%的引物编号、提取引物冗余对、冗余引物写成一行等6个步骤去除来源于自身部分同源序列以及与CMD释放的不同棉种相似性SSR引物,得到了非相似性引物,定名为CRIXXX (CRI即Cotton Research Institute)。并分别选用棉花12个种的代表性材料对其中100对进行引物功效评价,包括多态信息含量(polymorphism information content, PIC)及引物通用性研究。结果显示,从自主开发的100对SSR引物筛选出56对均能在12份材料间扩增出稳定明显的条带, 其中多态性引物35对,多态率占35%。引物的PIC变幅为0.097~0.888,平均为0.482;1对海岛棉EST-SSR引物在12份材料间的通用性为100%,25对亚洲棉引物通用性为81%,74对陆地棉引物通用性为80.1%。     

番薯属甘薯与牵牛EST-SSR标记开发及通用性分析

通过对番薯(Ipomoea)属的12 812条甘薯和28 422条牵牛EST唯一序列进行检索分析,在319条甘薯EST序列中发现了共328个EST-SSRs,平均每20.41 kb出现1个甘薯SSR;在936条牵牛EST序列中发现962个EST-SSRs,平均每17.87 kb出现1个SSR;在检索到的甘薯和牵牛EST-SSRs中,二核苷酸重复类型的SSR最多,分别占各自检索总数的44.51%和40.64%。利用SSR-ESTs分别设计了36对甘薯EST-SSR引物和92对牵牛EST-SSR引物,其中有28对甘薯引物在8个甘薯品种中有扩增产物,13对引物扩增产物具有多态性,多态性引物占所设计甘薯引物的36.1%;42对牵牛EST-SSR引物在甘薯基因组中可以扩增出清晰的条带,其中19对有表现较好的多态性,占设计的牵牛EST-SSR引物的20.7%。结果表明,牵牛EST-SSR标记在甘薯上具有通用性,可用于甘薯基因组多样性分析。     

三种瓜类作物SSR扩增的通用性与多态性分析

利用940对SSR引物,对甜瓜、黄瓜和西瓜三套重组自交系群体的作图亲本DNA进行PCR扩增,分析SSR引物在不同瓜类作物问的通用性与多态性.结果表明.上述SSR引物在甜瓜、黄瓜和西瓜等不同瓜类作物问具有一定的通用性,最小通用率超过24%,说明不同瓜类作物基因组同存在一定的相似性.但瓜类SSR引物的种属特异性很强,能够检测的多态率很低,三套作图亲本之间能够共用的多态性SSR引物比例甚至不足1%.     

枳壳EST-SSR标记的开发

GenBank上已公布的枳壳EST序列为开发新的SSR标记提供了宝贵的数据资源。本研究利用在线SSR鉴定软件SSRIT分析来自枳壳EST数据库的11029条Unigene序列。分析结果共发现327条EST序列含有348个SSR位点,占总数的2.96%。其中,三核苷酸重复的SSR类型最多,共有161个,占检索总数的46.26%。Primer 3.0设计合成58对EST-SSR引物,其中36对能扩增出产物,6对引物产生多态性分离,分别占所设计引物总数的62.07%和10.34%。本文研究成果为今后枳壳遗传多样性分析、遗传图谱构建及比较基因组等研究方面奠定了基础。     

鸟巢蕨EST-SSR标记的开发与应用

为加速分子标记在鸟巢蕨研究中的应用,利用鸟巢蕨EST序列开展了EST-SSR标记的开发和应用研究。利用MISA软件对鸟巢蕨EST序列进行SSR位点查找,分析EST-SSR的总体特征,并利用Primer 3.0软件设计40对引物,选用10份蕨类材料检测引物的多态性。从42 907条鸟巢蕨EST序列中检测到6 067个SSR位点,其出现频率为1/6.62 kb,包括106种重复基元。在鸟巢蕨EST-SSR中,二核苷酸重复(2 873个,47.35%)是最主要的类型,其次是三核苷酸(1 107个,18.25%)和单核苷酸(972个,16.02%)。出现最多的重复基元是AG/TC(1 371个,22.60%),其次是CA/GT(1 066个,17.57%)和A/T(628个,10.35%)。设计合成了40对EST-SSR引物对10份蕨类材料进行PCR扩增,26对引物能扩增出明显条带,其中17对引物扩增出了65条多态性条带,平均每对引物多态性带为3.82条。遗传分析表明,10份蕨类材料之间的遗传相似系数为0.338~0.815,可将供试蕨类材料分为2个类群。从鸟巢蕨EST序列中开发SSR标记是有效和可行的,开发的EST-SSR引物可用于蕨类植物遗传分析研究。     

海滨雀稗EST资源的SSR信息分析及EST-SSR标记开发

本研究探讨海滨雀稗EST序列中SSR位点的分布特征,开发EST-SSR引物分析其在草坪草种质资源遗传多样性研究中的应用。利用SSRIT软件对81220条海滨雀稗EST序列进行SSR位点搜索,分析EST-SSR的分布和特点,并利用Primer 3.0软件设计50对引物,选择10份草坪草对引物进行有效性和多态性检测。结果表明,从13705条海滨雀稗EST序列中检测到了22721个SSR位点,出现频率为16.87%。三核苷和单核苷酸重复为主要类型,所占比例分别为31.71%和29.28%。在检测到的163种基元中,出现最多的重复基元是A/T,其次是CCG/CGG和AGC/CTG。选择合成了50对EST-SSR引物,以海滨雀稗‘Adalady’基因组DNA为模板进行PCR扩增,其中37对引物能扩增出条带。利用这些引物对10份草坪草品种进行多态性检测,其中12对引物的扩增结果具有多态性。利用开发的EST-SSR标记对10份草坪草进行聚类分析,可将其分为4类。利用海滨雀稗EST序列开发SSR标记是可行的,开发的EST-SSR标记能有效用于草坪草遗传多样性研究。     

辣椒EST-SSR标记的开发

对4000条辣椒EST序列进行筛选,获得了119条含有SSR的EST,共设计出35对EST—SSR引物,占所有EST序列的0．87％。在所有的SSR—ESTs中,二核苷酸重复含量最高,其次是三核苷酸重复。GA和AGA分别是二,三核苷酸重复中含量最高的重复基元。以不育系（cytoplasmic male sterility,CMS）,保持系基因组DNA为模板,对EST—SSR引物进行筛选,首次获得Pe21、Pe26和Pe27等3对特异引物,并扩增出特异片段分别为CMSPe2160、CMSPe260、CMS Pe27300. EST—SSR标记作为功能基因的一部分,可以直接揭示或反映相关基因功能,特异标记基因的一部分序列,它的应用将对进一步研究辣椒胞质雄性不育分子机理带来一定的促进作用。     

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.     

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.     

Relation between wheat-rye crossability and seed set of common wheat after pollination with other species in the Hordeae

Summary Cultivars of common wheat (Triticum aestivum L. em. Thell.) of high wheat-rye (Secale cereale L.) crossability set more seed with pollen of other related species than did wheats of low wheat-rye crossability. This was found to be true for pollen parents from the genera Triticum, Aegilops, Secale, Agropyron and Elymus.     

Mechanisms and diversity of resistance to sorghum shoot fly,Atherigona soccata

Sorghum shoot fly, Atherigona soccata, is one of the important pests of postrainy season sorghums. Of the 90 sorghum genotypes evaluated for resistance to this pest, RHRB 12, ICSV 713, 25026, 93046 and 25027, IS 33844‐5, Giddi Maldandi and RVRT 3 exhibited resistance in postrainy season, while ICSB 463, Phule Anuradha, RHRB 19, Parbhani Moti, ICSV 705, PS 35805, IS 5480, 5622, 17726, 18368 and 34722, RVRT 1, ICSR 93031 and Dagidi Solapur showed resistance in rainy season, suggesting season‐specific expression of resistance to A. soccata. ICSB 461, ICSB 463, Phule Yasodha, M 35‐1, ICSV 700, 711, 25010, 25019 and 93089, IS 18662, Phule Vasudha, IS 18551 and 33844‐5 and Barsizoot had fewer deadhearts than plants with eggs across seasons, suggesting antibiosis as one of the resistance mechanism. Five genotypes exhibited resistance with high grain yield across seasons. Correlation, path and stepwise regression analyses indicated that leaf glossiness, seedling vigour, trichome density, oviposition and leaf sheath pigmentation were associated with the expression of resistance/susceptibility to shoot fly, and these can be used as marker traits to select and develop shoot fly‐resistant sorghums.     

Reaction of tritordeum to Fusarium culmorum and Septoria nodorum

Summary Hordeum chilense is a wild barley extensively used in wide crosses in the Triticeae. It could be a valuable source of resistance to Fusarium culmorum and Septoria nodorum. Some H. chilense x Triticum spp. amphiploids, named tritordeums, were more resistant than the parental wheat line to these diseases, others were not. Average contents of ergosterol and deoxynivalenol (DON) suggested that resistance to colonization by Fusarium was the highest for Hordeum chilense, followed by tritordeum and wheat in decreasing order. In particular, the H. chilense genotypes H7 and H17 enhanced the wheat resistance to F. culmorum in its tritordeum offsprings. Resistance to S. nodorum in tritordeum was not associated with tall plant height. There is sufficient genetic variation for resistance to F. culmorum and S. nodorum among tritordeum to allow the breeding of lines combining short straw and resistance to both diseases.     

Breeding apomictic bahiagrass (Paspalum notatum Flügge) with improved turf traits

The development of a turf‐type bahiagrass could have a remarkable impact on the billion‐dollar turf industry in the south‐eastern United States. The goals of this study were to (i) select bahiagrass genotypes with improved turf attributes and (ii) determine the reproductive mode and seed fertility of selected genotypes. The population included mutants, wild types, hybrids and standard cultivars evaluated at two locations for two years. Morphological variation was observed and clonal repeatability (H²) ranged from 0.27 to 0.90. Flowering was extremely reduced in some mutants, which also exhibited dark green colour, finer texture and reduced canopy height. Cytoembryological observations revealed that diploids reproduced sexually and tetraploids by obligate or facultative apomixis. Genotypes with superior seed fertility were identified, while others were comparable to the standard cultivars. While mutant plants exhibited great morphological variation, the random mutations did not affect their reproductive mode, validating the use of mutagenesis in apomictic species. The high H² indicated that selection for improved turf attributes could be successfully performed with the advantage of clonal seed propagation.     

Disease resistance in protected crops and mushrooms

Summary Cultivars of tomatoes, cucumbers, lettuce and peppers have been bred for resistance to one or more pathogens. Some tomato and cucumber cultivars have resistance to a wide range of diseases. Resistance has been transient in many cases and a succession of cultivars with new genes or new combinations of resistance genes has been necessary to maintain control. There has been a number of notable exceptions and these have included durable resistance to such pathogens asFulvia fulva and tomato mosaic virus. With lettuce the resistance situation is complicated by the occurrence of fungicide resistant pathotypes. There are no strains ofAgaricus bisporus purposely bred for disease resistance.In protected flower crops only resistance to Fusarium wilt in carnations has been purposely bred but differences in disease resistance are apparent in cultivars of many ornamental crops. This is particularly so in chrysanthemums where there are cultivars with resistance to many of the major pathogens. Similar situations occur with other flower crops and pot plants. Cultivars of some species have not been systematically investigated for resistance.The need for genetic resistance will increase with the further reduction, in the limits on pesticide use and an increasing public awareness and importance of pesticide pollution.ADAS is an executive agency of the Ministry of Agiculture, Fisheries and Food and the Welsh Office.     

Genetic constitution and diversity in four narrow endemic redwoods from the family Cupressaceae

The genetic constitution and diversity of four relictual redwoods are discussed in this review. These include monotypic genera of the family Cupressaceae: coast redwood (Sequoia sempervirens), giant sequoia (Sequoiadendron giganteum), dawn redwood (Metasequoia glyptostroboides), and alerce (Fitzroya cupressoides). All four species are narrow endemics, share a number of common phenotypic traits, including red wood, and are threatened species. Fossil history suggests that the ancestors of redwoods probably originated during the Cretaceous and Tertiary periods and flourished thereafter for millions of years. Towards the end of the Tertiary period began their decline and struggle for existence that continued during the subsequent geologic upheavals and climate changes, until the survival of the present-day redwoods in the current restricted locations in the world (USA, China, and South America). Although two species, Sequoiadendron and Metasequoia, are diploids (2n = 22), and the other two are polyploids: Fitzroya a tetraploid (2n = 4x = 44), and Sequoia a hexaploid (2n = 6x = 66); they all share the same basic chromosome number x = 11. The genome size in the hexaploid Sequoia is one of the largest (31,500 MB) in the conifers, while the genome sizes of diploid Metasequoia and Sequoiadendron are about one-third (~10,000 MB) of Sequoia. Genetic diversity in the redwoods is lower than most other gymnosperms, except in Sequoia, which seems to rank near the upper quarter of the coniferous forest trees. Genomic research is sparse in the redwoods, and should be pursued for a better understanding of their genome structure, function, and adaptive genetic diversity.     

Inheritance of fruit colour in a wild Russian red raspberry seedling

Summary A new gene, i, from a self-incompatible wild Russian Rubus idaeus seedling is described. The gene i interacts with the fruit colour gene T, Tii plants having apricot instead of red fruits, and spines, leaves and stems pigmented to varying degrees. A deficit of apricot-fruited seedlings in progenies segregating for i could be explained by linkage between i and the self-incompatibility S locus with a crossover value of approximately 26.7%.Segregations for spine colour in crosses and backcrosses of the Russian seedling with two green-spined (ttII) raspberries selected for their yellow fruit colour, showed that both the latter carried the spine colour intensifier gene P. This suggests that earlier authors were incorrect in attributing the apricot fruit colour developed by some tt plants to a pleiotropic effect of P.     

The induction in vitro of adventitious shoots in Rosa

Summary Adventitious shoots were formed on excised leaves, roots and callus of Rosa persica x xanthina and on excised leaves of R. laevigata and R. wichuraiana on culture media that included BAP and NAA as growth regulators. Shoots formed freely on freshly cultured callus of R. persica x xanthina but their production declined in successive cultures and ceased after twelve weeks. Transplantation to soil was improved by rooting plantlets in cellulose plugs in vitro and transferring plantlets to soil while still in the plugs.