Multivariate analysis of genetic variation in Aegilops tauschii from the world germplasm collection

A study of Aegilops tauschii subspecies constitution was undertaken. The data on allozyme and morphologic variation among 308 plants from 154 accessions were used for multivariate analysis. ACPH1 and (glume width)/(rachis segment width) ratio were found to be reliable criteria to distinguish between sspp. tauschii and strangulata.     

Oil and meal characteristics of core and non-core safflower accessions from the USDA collection

Germplasm evaluation of ex situ collections is needed to document collection characteristics, enhance utilization, and to determine collection needs. The objectives of this study were to (1) provide oil and meal evaluation information for a major portion of the United States Department of Agriculture (USDA) safflower (Carthamus tinctorius L.) collection, (2) compare ranges, variances and means between 203 core and 797 non-core accessions, and (3) determine if region of origin could be differentiated based on accession oil and meal characteristics. Means of the core and non-core accessions differed for % oil, palmitic acid, stearic acid, -tocopherols, and phenolic glucosides (both bitter and cathartic) (P<0.05). Differences between linoleic acid, oleic acid and -tocopherol means were not significant, and the variance between core and non-core accessions differed only for palmitic acid. Thus the core was not fully representative of the non-core accessions, but did capture a large fraction of the diversity in oil and meal factors present. Accessions from the Americas, China, South-West Asia, and South-Central Asia were differentiated using canonical discriminate analysis, but these regions overlapped to varying degrees with the E. European, Mediterranean, and E. African regions. Variation in % oil and fatty acids were generally more important than tocopherols and phenolic glucosides in differentiating accessions on a regional basis. The check cultivars Montola 2001 (high oleic) and Morlin (high linoleic) had oil and fatty acid content comparable to the maximums found in the collection. The ranges for tocopherols and phenolic glucosides among collection accessions, however, exceeded those of the cultivars, suggesting that collection accessions could be useful for genetic manipulation of these factors.     

Biomass production and related characters in the core collection of Pisum germplasm

Retention of residue on the soil surface following harvest is an effective method of reducing soil erosion from both wind and water. The pea crop produces small amounts of residue to effectively reduce soil erosion. Severe erosion occurs in pea production areas such as the Palouse Region of the US Pacific Northwest (PNW) when low residue crops such as spring pea or lentil are followed by fall-sown wheat. The current study was conducted to determine the range of total aboveground biomass (TAB), seed yield, and straw (residue) production from the plant identification (PI) accessions that comprise the core collection of Pisum germplasm. In addition, the potential for increasing seed yield and straw production simultaneously was evaluated. Three hundred and ninety PI accessions were screened in the field in 1996 and 1997. The variation for TAB, seed yield and straw production among the PI accessions exceeded that of the controls both years. Seed yield was positively correlated with straw production (r = 0.81, p< 0.01) indicating that seed and straw production can be increased simultaneously through positive selection for both traits. Significant favorable variation is present among accessions in the USDA core collection of Pisum germplasm which could be used to increase both seed yield and total biomass production of adapted breeding lines.     

Sampling strategy to develop a core collection of Uruguayan maize landraces based on morphological traits

Core collections were suggested to improve germplasmutilization. A core collection is a subset chosen to represent thediversity of a collection with a minimum of redundancies. Becausediversity is distributed between and within groups with differentdegrees of organization, an adequate classification of accessionsinto related groups should be performed prior to the selection of acore collection. Different classification strategies for theUruguayan Maize Collection were compared, and the best one was usedto select a core collection. The following classification strategieswere compared following a multivariate approach using the availablemaize data base: i) racial classification, ii) geographicorigin (south and north of the country), and iii) acombination of kernel type and geographic origin. The third optionwas considered the best classification rule, since it takes intoaccount two points which are closely related to the distribution ofdiversity: genotypic composition and geographic origin. The followingfive groups were identified in the collection: a) pop, b)floury, c) dent, d) southern flint-semiflints, ande) northern flint-semiflints. Eight core collections,each of 90 accessions, were selected, using different strategies toweight the groups in the core and to select the accessions from thegroups. The P, C, and L strategies were used and combined with eitherrandom selection within the group or the Relative Diversity method.Two samples of 90 accessions were obtained at random withoutconsidering the classification. The Relative Diversity methodcombined with the L strategy produced the best core collection, as itretained the highest percentage of the ranges for the 17 variablesincluded in the analysis. On average, 91% of the ranges wereretained in the core, confirming its representativeness.     

茶树黄酮醇合成酶基因的克隆与原核表达

本研究采用EST测序技术和RT—PCR技术,获得了一个茶树茶多酚代谢中的重要基因——黄酮醇合成酶（FLS）基因,在GenBank登录（GenBank accession No．EF205150）,其序列全长1317bp,其中开放阅读框长996bp,编码331个氨基酸,3]端有一个明显的多聚腺苷酸加尾信号,推测的蛋白分子量约为37．5kD,理论等电点为5．80。序列分析表明它与葡萄FLS基因序列的亲缘关系比较近。将该基因重组到表达载体pET-32a（＋）中进行原核表达,经IPTG诱导、SDS-PAGE检测,结果表明茶树黄酮醇合成酶基因能在大肠杆菌BL21中表达,电泳检测到一条大约61kD的外源蛋白,与预测的融合蛋白分子量相符。用Ni-NTA亲和层析柱对融合蛋白进行纯化,得到了纯度在90%以上的纯化蛋白,为进一步研究PET-FLS融合蛋白的活性及功能奠定了基础。     

Optimization of extraction of phenolic compounds from flax shives by pressurized low-polarity water

Pressurized low-polarity water (PLPW) extraction of phenolic compounds from flax shive was investigated using statistically based optimization and the "one-factor-at-a-time" method. Extraction variables examined using central composite design (CCD) included temperature, flow rate, and NaOH concentration of the extracting water. Extraction of phenolic compounds including p-hydroxybenzaldehyde, vanillic acid, syringic acid, vanillin, acetovanillone, and feruric acid was affected by temperature and NaOH concentration; and extraction of all phenolic compounds, except ferulic acid, increased with temperature and NaOH concentration of the extracting water. Flow rate had little effect on concentration of phenolic compounds at equilibrium, but the extraction rate at the early phase was higher for higher flow rates. The mechanism of PLPW extraction of flax shive phenolics was also investigated using a two-site kinetic model and a thermodynamic model. To determine the extraction mechanism, flow rate was varied from 0.3 to 4.0 mL/min while temperature and NaOH concentration were fixed at 180 degrees C and 0.47 M, respectively. The flow rate tests showed the extraction rates of total phenolic (TP) compounds increased with flow rate and can be described by a thermodynamic model. The results from the thermodynamic model demonstrated that a K(D) value of 30 agreed with the experimental data in the flow rate range of 0.3-4.0 mL/min. When the effect of the three independent variables was evaluated simultaneously using CCD, a maximum TP concentration of 5.8 g/kg of dry flax shive (DFS) was predicted from the combination of a high temperature (230.5 degrees C), a high initial concentration of NaOH (0.63 M), and a low flow rate (0.7 mL/min). Maximum TP concentration of 5.7 g/kg of DFS was obtained from extraction conditions of 180 degrees C, 0.3 or 0.5 mL/min, and 0.47 M NaOH at equilibrium. A second-order regression model generated by CCD predicted a maximum TP concentration of 5.8 g/kg of DFS under the same extraction conditions, which is well matched with the results from experimental data.     

An analysis of isozymic loci polymorphism in the core collection of the Polish Pisum genebank

The investigated material consisted of the core collection of the Polish Pisum gene bank at Wiatrowo and represented a described, monohybrid variation (type lines for characters/genes and their initial lines, tester lines with markers and lines from wild taxa, 266 accessions in total). A polymorphism in 18 isozymic loci was analyzed (Lap-1, Px-1, Aat-p, Aat-m, Aat-c, Pgm-c, Pgm-p, Gpi-c, Idh, Acp-1, Skdh, Est-1, Est-2, Est-3, Aldo, 6Pgd-p, 6Pgd-c, Dia-1). The practical outcome of the analysis was a selection of lines with a high number of fast or slow isozymic alleles, useful as tester lines with markers for chromosome mapping. A polymorphism was stated in all investigated loci with 2 to 4 alleles ranging from very fast, fast, slow to very slow. Fast and slow alleles occurred in all loci. In 11 loci the incidence of fast alleles was higher than slow ones. Conversly, slow alleles occurred more frequently than fast ones in 5 loci. Additionally, there was a low incidence of a very fast allele in 7 loci and of a very slow allele in 3 loci. A slow allele was rare in 2 loci. Primitive lines and land races were a source of these rare alleles. It is suggested to consider a frequent allele of a given locus found in a group of wild lines as a so-called wild type of Pisum.     

The development of a new approach for establishing a core collection using multivariate analyses with tulip as case

A methodology is developed for the establishment of a core collection based on phenotypic data. A case is worked out for the tulip cultivar collection of the Hortus Bulborum, consisting of approximately 1000 cultivars. The methodology is based on cluster analysis of phenetic characters, selection of cultivars from the resulting clusters according to a combination of the proportional and genetic diversity dependent strategies, optimisation of the selection, and validation by means of principal component analysis and by the diversity indices of Nei and Shannon & Weaver. A core collection with 104 cultivars (approx. 10%: CORE-1) did not give a sufficient representation of the existing diversity. A set of 152 cultivars (approx. 15%: CORE-2) showed a far better representation. The variation in resistance levels for Fusarium is almost completely represented in CORE-2. Although a set of 200 cultivars (approx. 20%: CORE-3) reached a nearly complete representation of the total diversity, a better representation of the resistance levels was not achieved. The newly presented methodology for defining a core collection appears to be a useful approach. The included steps for optimising and validating the choice of accessions makes it a reliable and broadly applicable method.     

The principal component scoring: A new method of constituting a core collection using quantitative data

The principal aims of a core collection are: 1) to include the maximum of diversity of the base collection in a sample of minimum size; and 2) to avoid redundancies. We present here a new method (the Principal Component Scoring) which fulfills these aims. The use of P.C.S. has consequences for sampling stratification and choice of sample size. P.C.S. requires quantitative data, but with small changes can be used for qualitative data.     

拟南芥AtHOS10基因的克隆及其表达载体的构建

AtHOS10基因对植物冷调节起到重要作用,可以通过控制ABA合成来改变植物的脱水胁迫耐性。本文利用PCR方法从Columbia生态型的拟南芥基因组中扩增出冷调节基因AtHOS10,并插入克隆载体。序列分析表明,克隆片段长3453bp,与目前发表的序列完全一致。将pUC-AtHOS10和pBI-HEM进行酶切重组构建表达载体pBI-AtHOS10,然后将表达载体转化导入农杆菌中,为侵染烟草作准备。     

A method for establishing a core collection of Saccharum officinarum L. germplasm based on quantitative-morphological data

A technique is proposed for establishing a representative core collection of S. officinarum accessions from the world collection of sugarcane germplasm maintained at the Sugarcane Breeding Institute-Research Centre, Cannanore, India. In the proposed method, the accessions were first sorted based on their relative contributions to the total variability by means of principal component scores based on a set of quantitative characters. Then, the cumulative proportion of their contributions to the total variance was computed. A logistic regression model was fitted to evaluate the functional relationship between the cumulative proportion of variance and the number of accessions. The size of the core set was decided as the inflection point on that fitted curve, i.e., the point beyond which the rate of increase in cumulative proportion of variability contributed by an accession began to decline. A method for eliminating entries with a high degree of similarity from the selected core set is also proposed.     

Analysis of duplication in the Spanish durum wheat collection maintained in the CRF-INIA on the basis of agro-morphological traits and gliadin proteins

A total of 106 potential duplicate cases involved 277 accessions were detected on the basis of passport data in the durum wheat collection maintained in the CRF-INIA. Similarity between accessions was measured by agro-morphological traits. The 90% of the agro-morphological duplication were verified with gliadin proteins, allowing identification of similar material with greater refinement than agro-morphological data. However, the results indicated not to decide for rationalisation only on the basis of molecular data.     

Variation in flowering time in sorghum core collection and mapping of QTLs controlling flowering time by association analysis

A wide range of variation in flowering time was observed within a diversity research set of 107 sorghum accessions ranging from 56 to 133 days. Accessions were classified into early medium and late flowering groups. 45 accessions were grown under three different environments of photoperiod (11, 12 and 15 h). Sorghum accessions gradually responded to the decreasing of day-length. The 12 h of photoperiod could be considered as a threshold above which day-length delays the flowering time in sorghum. Association analysis was performed to identify the QTLs controlling flowering time and photoperiod sensitivity using 107 accessions of sorghum grown under natural condition and 45 accessions grown under controlled conditions. Four QTLs controlling flowering time were detected under natural condition of day-length at threshold 2.5 using K model. A total of seven flowering time loci were detected under controlled conditions of day-length. One QTL controlling photoperiod sensitivity was detected on chromosome 1 and one QTL controlling photoperiod insensitivity was detected on chromosome 4.     

芥菜新型几丁质酶基因在酵母(Pichia pastoris)中的分泌表达

BjCHI1基因是从芥菜（Brassica juncea)中克隆的1个新型几个质酶基因，编码含2个几丁质结合域的几丁质酶，本研究曾试图在大肠杆菌中表达该基因而未成功，因此试在酵母中表达，按照BjCHI1cDNA全序列设计特异引物，通过PCR扩增获得成熟肽第1密码子至最后密码子之间的片段，克隆到载体pPIC9K上，测序验证无误后，转化酵母KM71，获得6个抗1.0mg/mL遗传霉素（G418）水平的阳性转化子，PCR鉴定表明外源基因已经整合到酵母染色体上，此6个菌株经初步诱导，均表达出预期的蛋白质，对其中表达效率相对较高的KM71-17-1菌株作进一步的诱导表达，结果表明在诱导第3天后产生的外源蛋白达最大值，至此，成功地获得了BjCHI1蛋白，为进一步研究奠定了基础。