构树种质资源遗传多样性SRAP分子标记研究

构树（Broussonetia papyrifera L.）为桑科（Moraceae）构树属（Broussonetia）落叶乔木，广泛分布于亚洲东部及太平洋岛屿。它是我国重要的经济林木，具有重要的经济价值，其树皮纤维品质优良，自古就是造纸的优良原料；叶片可用作饲料；果实具有重要的药用价值；环境适应性强，是迅速绿化荒山、荒滩和盐碱地的理想树种。因此对构树这些特性的深入研究和开发利用具有非常重要的实际应用价值。本研究以日本和国内构树主要分布区域的的10种生态型及杂交构树共23份材料，摸索并改进了构树DNA的提取方法，建立了稳定的SRAP分子标记体系，以杂交构树组培苗为材料从120对引物组合中筛选出条带信息较高的17对SRAP引物，以这些引物对23份构树材料进行PCR扩增和标记分析。 本研究取得的主要结果如下： （1）本实验首次将SRAP技术应用于构树的研究中，建立起构树稳定的SRAP-PCR反应体系；实验中对影响扩增的5个主要因素进行了优化，确定20 μL PCR反应体系中各因素最适浓度：模板DNA浓度80 ng/(20 μL)，Mg2+浓度2.0 mM，dNTP浓度0.6 mM，引物浓度0.8 mM，Taq 酶浓度1.5 U/(20 μL)。 （2）从120对引物组合中筛选出来的17对SRAP引物对21份不同生态型构树样本（21份材料指的是除两份杂交构树材料外的其它生态型构树，以下同）进行PCR扩增，共扩增出439条带，平均每对引物25.5条，其大小介于100～1,000 bp之间，其中多态性条带319条，占总数的72.67%。 （3）用Popgene1.32软件进行分析，计算出Shannon信息指数（I）值为0.2275（0.2042），物种水平的Nei基因多样性（H）值为0.1336（0.1436），表明各生态型构树之间的平均遗传多态性不高，中国大陆各生态型构树Shannon信息指数（I）值仅为0.1675（0.2271），物种水平的Nei基因多样性（H）值为0.1039（0.1540），群体遗传多样性较低，构树的遗传分化主要存在于中国和日本之间。 （4）用NTSYS-2.10e软件进行聚类分析，发现不同生态型构树按距离关系远近及分布区域可划分为不同类群。在遗传相似系数0.57附近，对21份材料进行聚类分析发现其可分为两个群体，一类为日本生态型，另一类为中国生态型，表明日本构树与中国野生种构树种源遗传相似性较小。中国各生态型构树在遗传相似系数0.91处可分为5类，总体而言，中国各地区之间的构树遗传相似度较高。对杂交构树分析表明，其亲缘关系与日本构树（母本）更接近。 （5）日本及杂交构树的SCAR标记。本研究找到两条日本及杂交构树的特异性条带，回收、测序，再根据序列往里重新设计引物，转变成稳定性更好，更直观的SCAR标记，这为挑选性状优良的日本及杂交构树提供重要的参考，对其育种有一定的指导意义。其中一条片段经与NCBI数据库比对发现与拟南芥磺基转移酶家族基因具有较高的同源性。

紫薇种质资源的研究

本文回顾了紫薇在国内外的栽培历史。在中国，紫薇的栽培虽历史悠久，但种质资源缺少系统的研究。本研究通过宏观与微观的技术和方法，探索紫薇种间及品种间的演化关系，并为今后开展种质鉴定、 品种分类以及有计划、有目的的进行新品种的培育提供依据，以期达到紫薇种质资源多样性保护的目的。主要的结果及结论： 1．认为紫薇栽培的历史是紫薇不同种质进行组合和渗透的历史，也在一定程度上反映了品种演化的历史。并提出除了抗病、矮型紫薇的育种外，还要重视重瓣紫薇的育种。 2．通过对紫薇的形态学性状进行聚类分析，寻找到了评价紫薇种质资源的标准，进而提出栽培观赏植物种质资源的评价标准， 即将其性状分为种源性状和观赏性状，这种对观赏植物性状的分解反映了栽培植物不同于野生植物的特点。同时，对其形态学性状进行主分量分析表明，花色与其它营养器官性状相关性不大，很难从其营养器官性状推测花的颜色。 3．通过对紫薇的分子生物学研究，建立了紫薇的DNA提取流程及RAPD扩增体系，利用RAPD标记可以进行紫薇种质的鉴定及探求不同种质间的关系。 4．综合形态学及分子生物学的结论，建立了紫薇种质资源的划分体系，在此基础上，建成了种质资源圃及紫薇种质资源数据库。

小麦—偏凸山羊草6Mv/6B代换系的研究

小麦条锈病（Puccinia striiformis f. sp. tritici）是世界性小麦病害，可导致受害小麦减产30%以上，甚至绝收。小麦条锈病在我国西南、华北麦区危害严重，四川麦区是小麦条锈病发病最重的地区之一，每年因条锈病流行造成小麦产量损失巨大。利用抗条锈病品种是控制该病害最安全、经济的有效途径，因此挖掘利用抗病新基因，开展抗病遗传基础研究是当前育种工作中面临的重要任务。 偏凸山羊草（Aegilops ventricosa，DDMvMv，2n=28）是一年生草本植物，起源于地中海西部沿岸地区，具有对小麦白粉病、锈病等高抗或免疫、耐盐、抗寒、蛋白质含量高等优良性状，是小麦遗传育种很好的种质资源。本研究以高抗条锈病的小麦—偏凸山羊草6Mv/6B代换系（Moisson 6Mv/6B）为材料，对其含有的带条锈病抗性基因的偏凸山羊草6Mv染色体在四川小麦背景中的传递情况、与小麦—簇毛麦双端体附加系所具有的白粉病抗性的聚合以及对Moisson 6Mv/6B进行电离辐射诱变筛选抗条锈病的小麦—偏凸山羊草易位系三个方面进行了研究。取得的主要研究结果如下： 1. Moisson 6Mv/6B与高感条锈病的四川地区普通小麦品种绵阳26、绵阳93-124和SW3243的杂种F1与其普通小麦亲本分别作为父、母本回交，通过对其BC1和F2的结实率、根尖细胞有丝分裂中期染色体的观察以及对条锈病抗性的鉴定，发现含6Mv染色体的F1植株作母本时的回交结实率（83.10%）普遍高于含6Mv染色体的F1植株作父本（48.61%），结实率与普通小麦基因型密切相关（χ2=34.15>>χ20.05=5.99（df=2））；6Mv染色体在三种四川小麦中通过雌、雄配子传递的传递方式与其传递率间没有显著相关性，其传递率与普通小麦基因型呈显著相关性（χ2=6.42>χ20.05=5.99（df=2））。 2. Moisson 6Mv/6B与高抗白粉病的小麦—簇毛麦双端体附加系Pana（2n=42+2t）正反杂交，希望在聚合两者抗性的同时观察不同受体背景下的抗性反应。对Moisson 6Mv/6B和Pana正反杂交的结实率、杂交后代的农艺性状进行观察，并对杂交后代进行基因组荧光原位杂交（GISH）分析及条锈病和白粉病的抗性鉴定。结果表明Moisson 6Mv/6B作母本时杂交结实率（80.56%）高于Pana作母本时（58.33%），结实率与杂交方式间紧密相关（χ2=4.96>χ20.05=3.84（df=1））；Moisson 6Mv/6B和Pana杂交后代株高比最高亲本高约10cm，成熟期也较两亲本提前两个星期左右；正反杂交后代中具有偏凸山羊草6Mv染色体的植株具有条锈病抗性，具有簇毛麦端体的植株具有白粉病抗性，同时筛选到4株含有偏凸山羊草和簇毛麦遗传物质并对条锈病和白粉病兼抗的材料，证明来自偏凸山羊草6Mv染色体的条锈病抗性与来自簇毛麦端体的白粉病抗性已经聚合在一起，且没有产生相互抑制的作用，暗示通过这两个抗性基因的聚合是完全能获得兼抗条锈病和白粉病的小麦新种质。 3. 对Moisson 6Mv/6B在减数分裂时期的成株进行总剂量为6Gy、辐射频率为120rad/min的60Co-γ射线辐射，对辐射植株自交后代进行农艺性状及根尖细胞有丝分裂中期染色体形态观察和条锈病抗性鉴定。结果为辐射植株自交结实率为2.22%，根尖细胞有丝分裂中期的染色体存在明显碎片，辐射自交后代植株对条锈病具有成株期抗性。 小麦—偏凸山羊草6Mv/6B代换系对条锈病抗性稳定，是培育条锈病抗性品种的良好供体。本研究证明在四川小麦背景中要利用该品种抗性，在结实数满足需要时，可将其作父本，亦可作母本，但关键是要选择好一个优良的受体基因型；同时其条锈病抗性与来自簇毛麦的白粉病抗性没有相互抑制作用，可将两者抗性有效聚合用于小麦育种中。 Wheat stripe rust (Puccinia striiformis f. sp. Tritici) is a worldwide disease of wheat, and could lead to victims of 30 percent or even total destruction of wheat production. Wheat stripe rust harms badly in China's southwest and North China. Sichuan province is one of the regions damaged by wheat stripe rust heavily. The use of resistant varieties is the most secure and economical way to control the wheat stripe rust. Therefore, it is essential to identify new disease-resistant genes and genetically research of disease resistance. Aegilops ventricosa (DDMvMv, 2n = 28) is an annual herbaceous plant, originating in the coastal areas of the western Mediterranean, with good characters such as resistance of wheat powdery, rust, salt, cold and high protein content. It is a good germplasm resource. In this study, the wheat- Aegilops ventricosa 6Mv/6B substitution line Moisson 6Mv/6B (highly resistant to the wheat stripe rust) was used to study on the transmission of chromosome 6Mv of Aegilops ventricosa in different genetic background of Sichuan wheat varieties, hybridization with wheat- Haynaldia villosa ditelosomic addition line Pana (highly resistant to the powdery mildew) and screening of wheat- Aegilops ventricosa translocation line by exposuring Moisson 6Mv/6B under ionizing radiation. The main results are as following: 1. Moisson 6Mv/6B was crossed with Sichuan wheat varieties mianyang26, mianyang93-124 and SW3243 (highly susceptible to stripe rust), respectively. Their F1 hybrids were further backcrossed as male and female to corresponding wheat varieties. The seed-setting rate, chromosomes confirmation in the mitotic metaphase of root tip cells, and resistance to stripe rust of the subsequent BC1 and F2 plants were investigated. The average seed-setting rate of backcross via 6Mv as female donor (83.10%) was higher than that of backcross via 6Mv as male donor (48.61%), suggesting that the seed-setting rate was associated with the wheat genotypes（χ2=34.15>>χ20.05=5.99（df=2））. In all analyzed populations, transmission frequencies of chromosome 6Mv were not correlated with the ways of 6Mv through male or through female. However, transmission frequencies of chromosome 6Mv were significantly correlated with Sichuan wheat genotypes（χ2=6.42>χ20.05=5.99（df=2））. 2. To aggregating the resistances to stripe rust and powdery mildew, as well as research on the resistance reactions in different genetic background, Moisson 6Mv/6B was reciprocally hybrided with the wheat- Haynaldia villosa ditelosomic addition line Pana (highly resistant to the powdery mildew). The seed-setting rate, agronomic characters, genomic in situ hybridization (GISH) of hybrid progenies，and resistances to stripe rust and powdery mildew were investigated. The results showed that the seed-setting rate of hybridization via Moisson 6Mv/6B as female donor (80.56%) was significant higher than that via Pana as female donor (58.33%). The seed-setting rate was associated with the hybrid methods (χ2 = 4.96> χ20.05 = 3.84 (df = 1)). The plant height of hybrid progenies was about 10 cm higher than Pana, the parent with maximum height. And the maturity of hybrid progenies was about two weeks earlier than that of the parents. In the hybrid progenies, the plants with the 6Mv chromosome have the resistance to stripe rust and the plants with the telosome from Haynaldia villosa have the resistance to powdery mildew. It was found that four plants with both the 6Mv chromosome and the telosome from Haynaldia villosa were resistant to stripe rust and powdery mildew. It indicated that the resistance to stripe rust and powdery mildew aggregated, and no mutual inhibition was found. It implied that the aggregation of the two resistance genes was able to provide the new wheat germplasm with the resistances to stripe rust and powdery mildew. 3. Moisson 6Mv/6B was irradiated with 60Co-γ rays of 6Gy (120rad/min) during meiosis. The agronomic characters and chromosomes confirmation in the mitotic metaphase of root tip cells，as well as resistance to stripe rust were investigated. The seed-setting rate of irradiated plants was only 2.22%. The chromosomes in mitotic metaphase had clear fragments. The resistance to stripe rust of progeny of irradiated plants was the adult-plant resistance. The wheat- Aegilops ventricosa 6Mv/6B substitution line is a good stripe rust resistance donor for its stabile resistance. Our study demonstrated that the key for use the resistance is to choose a good receptor. There is no difference between Moisson 6Mv/6B be the female and be the male if the seed number meets the requirement. At the same time, the stripe rust resistance of Moisson 6Mv/6B did not have the mutual inhibition with the powdery mildew resistance from Haynaldia villosa. It is able to aggregate the two resistances for wheat breeding.

Variation among Australian accessions of the wild mungbean (Vigna radiata ssp sublobata) for traits of agronomic, adaptive, or taxonomic interest

The wild mungbean, Vigna radiata ssp. sublobata, is an 'old world' tropical species indigenous throughout the better watered areas of northern Australia. Variation among 115 accessions, mainly from Australia, West Timor, and Papua New Guinea, was evaluated for several diverse traits. The plants were cultivated in the field at 2 sowing dates, at both a tropical and a subtropical location, with 6 accessions from India and a mungbean cultivar for comparison. Substantial variation was identified for traits of potential agronomic, adaptive, or taxonomic interest. For some traits, like phenology, the variation appeared to be systematic, with plausible underlying physiological and/or adaptive explanation. Among accessions, wild type traits, like prostrate habit, more gracile morphology, twining form, and small hard seeds, tended to be associated. There was a general geographic trend for lines collected from locations more remote from where mungbean has historically been cultivated to show greater expression of wild type traits, with few 'traits of domestication' evident in the Australian accessions. Some of the identified variation, e. g. higher seed protein content, hardseededness, and putative disease resistance, may be of value in mungbean variety improvement. A more targetted evaluation of the collection would likely reveal other adaptations, especially tolerance to environmental stresses. As such, the wild accessions are a potentially valuable if under-utilised germplasm resource.

Patterns of diversity in fatty acid composition in the Australian groundnut germplasm collection

Knowledge of the amounts and types of fatty acids in groundnut oil is beneficial, particularly from a nutritional standpoint. Germplasm evaluation data for fatty acid composition on 819 accessions of groundnut (Arachis hypogaea L.) from the Australian Tropical Field Crops Genetic Resource Centre, Biloela, Queensland were examined. Data for eight quantitative fatty acid descriptors have been documented. Statistical assessment, via methods of pattern analysis, summarised and described the patterns of variation in fatty acid composition of the groundnut accessions in the Australian germplasm collection. Presentation of the results from principal components analysis and hierarchical cluster analysis using a biplot was shown to be a very useful interpretative tool. Such a biplot enables a simultaneous examination of the relationships among all the accessions and the fatty acids. Unlike that information available via database searches, the results from contribution analysis together with the biplot provide a global picture of the diversity available for use in plant breeding programs. The use of standardised data for eight fatty acids, compared to using three specific fatty acids, provided a better description of the total diversity available because it remains relevant with possible changes in the nutritional preferences for fatty acids. Fatty acid composition was found to vary in relation to the branching pattern of the accessions. This pattern is generally indicative of the botanical types of groundnuts; Virginia (alternate) compared to Spanish and Valencia (sequential) botanical types.

Mixed data types and the use of pattern analysis on the Australian groundnut germplasm data

Data in germplasm collections contain a mixture of data types; binary, multistate and quantitative. Given the multivariate nature of these data, the pattern analysis methods of classification and ordination have been identified as suitable techniques for statistically evaluating the available diversity. The proximity (or resemblance) measure, which is in part the basis of the complementary nature of classification and ordination techniques, is often specific to particular data types. The use of a combined resemblance matrix has an advantage over data type specific proximity measures. This measure accommodates the different data types without manipulating them to be of a specific type. Descriptors are partitioned into their data types and an appropriate proximity measure is used on each. The separate proximity matrices, after range standardisation, are added as a weighted average and the combined resemblance matrix is then used for classification and ordination. Germplasm evaluation data for 831 accessions of groundnut (Arachis hypogaea L.) from the Australian Tropical Field Crops Genetic Resource Centre, Biloela, Queensland were examined. Data for four binary, five ordered multistate and seven quantitative descriptors have been documented. The interpretative value of different weightings - equal and unequal weighting of data types to obtain a combined resemblance matrix - was investigated by using principal co-ordinate analysis (ordination) and hierarchical cluster analysis. Equal weighting of data types was found to be more valuable for these data as the results provided a greater insight into the patterns of variability available in the Australian groundnut germplasm collection. The complementary nature of pattern analysis techniques enables plant breeders to identify relevant accessions in relation to the descriptors which distinguish amongst them. This additional information may provide plant breeders with a more defined entry point into the germplasm collection for identifying sources of variability for their plant improvement program, thus improving the utilisation of germplasm resources.

Pattern analysis of the diversity of morphological plant attributes and herbage yield in a world collection of white clover (Trifolium repens L.) germplasm characterised in a summer moisture stress environment of Australia

Information on the variation available for different plant attributes has enabled germplasm collections to be effectively utilised in plant breeding. A world sourced collection of white clover germplasm has been developed at the White Clover Resource Centre at Glen Innes, New South Wales. This collection of 439 accessions was characterised under field conditions as a preliminary study of the genotypic variation for morphological attributes; stolon density, stolon branching, number of nodes. number of rooted nodes, stolon thickness, internode length, leaf length, plant height and plant spread, together with seasonal herbage yield. Characterisation was conducted on different batches of germplasm (subsets of accessions taken from the complete collection) over a period of five years. Inclusion of two check cultivars, Haifa and Huia, in each batch enabled adjustment of the characterisation data for year effects and attribute-by-year interaction effects. The component of variance for seasonal herbage yield among batches was large relative to that for accessions. Accession-by-experiment and accession-by-season interactions for herbage yield were not detected. Accession mean repeatability for herbage yield across seasons was intermediate (0.453). The components of genotypic variance among accessions for all attributes, except plant height, were larger than their respective standard errors. The estimates of accession mean repeatability for the attributes ranged from low (0.277 for plant height) to intermediate (0.544 for internode length). Multivariate techniques of clustering and ordination were used to investigate the diversity present among the accessions in the collection. Both cluster analysis and principal component analysis suggested that seven groups of accessions existed. It was also proposed from the pattern analysis results that accessions from a group characterised by large leaves, tall plants and thick stolons could be crossed with accessions from a group that had above average stolon density and stolon branching. This material could produce breeding populations to be used in recurrent selection for the development of white clover cultivars for dryland summer moisture stress environments in Australia. The germplasm collection was also found to be deficient in genotypes with high stolon density, high number of branches high number of rooted nodes and large leaves. This warrants addition of new germplasm accessions possessing these characteristics to the present germplasm collection.

Highly heritable resistance to root-lesion nematode (Pratylenchus thornei) in Australian chickpea germplasm observed using an optimised glasshouse method and multi-environment trial analysis

Pratylenchus thornei is a root-lesion nematode (RLN) of economic significance in the grain growing regions of Australia. Chickpea (Cicer arietinum) is a significant legume crop grown throughout these regions, but previous testing found most cultivars were susceptible to P. thornei. Therefore, improved resistance to P. thornei is an important objective of the Australian chickpea breeding program. A glasshouse method was developed to assess resistance of chickpea lines to P. thornei, which requires relatively low labour and resource input, and hence is suited to routine adoption within a breeding program. Using this method, good differentiation of chickpea cultivars for P. thornei resistance was measured after 12 weeks. Nematode multiplication was higher for all genotypes than the unplanted control, but of the 47 cultivars and breeding lines tested, 17 exhibited partial resistance, allowing less than two fold multiplication. The relative differences in resistance identified using this method were highly heritable (0.69) and were validated against P. thornei data from seven field trials using a multi-environment trial analysis. Genetic correlations for cultivar resistance between the glasshouse and six of the field trials were high (>0.73). These results demonstrate that resistance to P. thornei in chickpea is highly heritable and can be effectively selected in a limited set of environments. The improved resistance found in a number of the newer chickpea cultivars tested shows that some advances have been made in the P. thornei resistance of Australian chickpea cultivars, and that further targeted breeding and selection should provide incremental improvements.

Bionomics, Resource Characteristics And Distribution Of The Threatened Freshwater Fishes Of Kerala

The previous faunistic studies were concentrated.on the taxonomical and zoogeo— graphical aspects. These studies contributed to many new additions to the fish fauna of Kerala meanwhile many species described earlier are reported missing in recent years. Many fish species were collected only once or twice by scientists. Detailed information on distribution, habitat, feeding habits, reproduction, population size, etc. are available only with regard to a very few fish species. A meaningful assessment on the biodiversity status of the majority of freshwater fishes cannot be done for want of sufficient data base and therefore, no suitable conservation and management programmes are forthcoming for the protection and preservation of the unique fish germplasm resources of Kerala. The present study was conceptualised and undertaken mostly aiming at bridging these gaps by generating an authentic data base on the distribution, resource characteristics and bionomics of the threatened fishes inhabiting the rivers of Kerala. Osteobrama bakeri (Day) is an endemic fish having a very highly restricted and fragmented distribution in Periyar, Chalakudy, Kabini, Kallada and Meenachil rivers of Kerala. This belongs to vulnerable category and is locally known as Mullanpaval which is valued as food fish. Besides, due to its vibrant and attractive colouration and easiness for domestication, it has great potential for being propagated as an ornamental fish. Hitherto, no information is available on the bionomics and resource characteristics of this species. Studies on detailed life history traits are indispensable for fishery management, development of captive breeding technique and implementation of various conservation programmes. In the present study, a pioneer attempt is also made to investigate the life history traits, resource characteristics, proximate composition, etc. of O.baken'.

Grain yield components of pearl millet under optimum conditions can be used to identify germplasm with adaptation to and zones

There is evidence that high-tillering, small-panicled pearl millet landraces are better adapted to the severe, unpredictable drought stress of the and zones of NW India than are low-tillering, large-panicled modern varieties, which significantly outyield the landraces under favourable conditions. In this paper, we analyse the relationship of and zone adaptation with the expression, under optimum conditions, of yield components that determine either the potential sink size or the ability to realise this potential. The objective is to test whether selection under optimal conditions for yield components can identify germplasm with adaptation to and zones in NW India, as this could potentially improve the efficiency of pearl millet improvement programs targeting and zones. We use data from an evaluation of over 100 landraces from NW India, conducted for two seasons under both severely drought-stressed and favourable conditions in northwest and south India. Trial average grain yields ranged from 14 g m(-2) to 182 g m(-2). The landraces were grouped into clusters, based on their phenology and yield components as measured under well-watered conditions in south India. In environments without pre-flowering drought stress, tillering type had no effect on potential sink size, but low-tillering, large-panicled landraces yielded significantly more grain, as they were better able to realise their potential sink size. By contrast, in two low-yielding and zone environments which experienced pre-anthesis drought stress, low-fillering, large-panicled landraces yielded significantly less grain than high-tillering ones with comparable phenology, because of both a reduced potential sink size and a reduced ability to realise this potential. The results indicate that the high grain yield of low-tillering, large-panicled landraces under favourable conditions is due to improved partitioning, rather than resource capture. However, under severe stress with restricted assimilate supply, high-tillering, small-panicled landraces are better able to produce a reproductive sink than are large-panicled ones. Selection under optimum conditions for yield components representing a resource allocation pattern favouring high yield under severe drought stress, combined with a capability to increase grain yield if assimilates are available, was more effective than direct selection for grain yield in identifying germplasm adapted to and zones. Incorporating such selection in early generations of variety testing could reduce the reliance on random stress environments. This should improve the efficiency of millet breeding programs targeting and zones. (c) 2005 Elsevier B.V. All rights reserved.

Molecular characterization of accessions of a rare genetic resource: sugary cassava (Manihot esculenta Crantz) from Brazilian Amazon

2016