无核葡萄种质亲缘关系的SRAP研究

为充分了解及更合理地利用无核葡萄的种质资源,辅助无核葡萄育种,本研究采用SRAP分子标记技术,对23 个国内外主流无核葡萄品种的种质亲缘关系进行分析。结果表明：从45 对SRAP引物中筛选出38 对多态性引物,共扩增出236 条谱带,其中171 条为多态性谱带,多态性谱带百分率为72.4%。遗传相似分析显示,供试材料间遗传相似系数的变化范围为0.58~0.90,遗传距离在1.0313~4.9643 范围内变化。在遗传相似系数为0.61 处,可将供试材料分为2 个大类群,在相似系数为0.66 处,Ⅰ类群和Ⅱ类群均可分为2个亚群。说明23个品种总体来讲亲缘关系较远,特别欧美杂种具有较高的遗传多样性,部分欧亚种品种间亲缘关系较近,杂交育种时应注意亲本选择。     

黄化病对葡萄生长发育的影响

[目的]为了研究黄化病对葡萄生长发育的影响。[方法]以吐鲁番主栽品种无核白和无核白鸡心为试材,通过对不同黄化程度葡萄的叶片生长量、叶绿素含量及果实产量等指标进行对比分析。[结果]黄化对葡萄植株生长、果实产量和品质造成了严重危害。随着葡萄黄化程度的加剧,两种葡萄植株的单叶重、叶长、叶宽及单叶厚等逐渐减小,且黄化株生长量较正常株明显下降。黄化植株葡萄的产量较低,两种葡萄正常园产量分别是黄化园产量的2.47倍和2.51倍,果粒大小和可溶性固形物也显著低于正常植株。采用不同方法提取叶绿素含量比较得出,利用丙酮研磨法提取两种葡萄叶绿素含量的效率均优于乙醇浸泡法。[结论]黄化病造成葡萄叶片光合代谢失调,植株生长迟缓,严重威胁着吐鲁番葡萄产业的健康发展。对黄化病的发生原因进行深入分析,并采取有效的防治措施是现在葡萄生产上比较棘手的问题。     

梁山慈竹高通量转录组测序及差异表达基因分析

分析梁山慈竹及其体细胞突变体的转录组,挖掘功能基因并对其差异表达基因进行筛选和分析,为梁山慈竹遗传改良提供理论依据。利用RNA-Seq技术进行转录组测序,对测序结果进行de novo拼接和功能注释;并对差异表达基因进行筛选及COG、GO、KEGG数据库中进行比对注释,此外,基于Swiss-Prot功能注释结果,分析纤维素和木质素相关功能基因的表达量差异。测序结果表明,共获得86 575 631条reads,de novo组装得到84 741条unigenes,共有49 829条被Nr、COG、GO、KEGG、Swiss-Prot注释。从梁山慈竹实生植株(对照)和体细胞突变体No.30这2个测序样本中,筛选出3 572条差异表达unigenes,757条差异表达unigenes在COG分类体系中具有详细的蛋白功能释义,2 213条差异表达unigenes在GO数据库具有功能定义,385条unigenes被注释到94条KEGG Pathways中。纤维素合成相关纤维素合酶、过氧化物酶、泛素连接酶和热休克蛋白在梁山慈竹体细胞突变体No.30中表达量升高,木质素合成相关MYB4、4-香豆酸CoA连接酶、肉桂醇脱氢酶、肉桂酰-CoA还原酶和漆酶在突变体中表达量降低。提供了全面的梁山慈竹转录组信息,获得了一批在梁山慈竹纤维素和木质素生物合成过程中有重要功能的基因序列。     

无核葡萄胚败育机制研究进展

为了更加全面地掌握无核葡萄胚败育机制研究进展,明晰相关的研究结果和存在问题。笔者对 国内外近年来相关的研究成果进行了综述,发现研究成果主要集中在细胞学、生理生化、无核性状遗传 规律、无核基因的分子标记和无核相关基因功能5 个方面。然而有关胚发育及败育过程中基因种类、基 因间如何互作,调控网络等方面研究还相对欠缺。最后对今后的趋势提出展望,认为可运用多组学联合 分析从整体上探讨无核葡萄胚败育机制。     

Genetic analysis of flowering and maturity time in high latitude spring wheat

Due to the short growing season in the high northern latitudes, the development of early maturing spring wheat (Triticum aestivum L.) cultivars is important to avoid frost damage which can lower production and quality. We investigated earliness of flowering and maturity, and some associated agronomic traits, using a set of randomly selected high northern latitude adapted spring wheat cultivars (differing in maturity) and their F₁ and F₂ crosses made in a one-way diallel mating design. The parents, and their F₁ and F₂ crosses were evaluated under field conditions over 2 years. Anthesis and maturity times were controlled by both vernalization response and earliness per se genes, mainly acting additively. Non-additive genetic effects were more important in controlling grain fill duration, grain yield and plant height. Additive × additive epistatic effects were detected for all traits studied except time to anthesis. Segregation analyses of the F₂ populations for time to anthesis indicated the presence of different vernalization response genes. Molecular genetic analyses revealed the presence of Vrn-A1 and Vrn-B1 genes in the parental cultivars. Narrow-sense heritability was medium to high (60–86%) for anthesis and maturity times but low to medium (13–55%) for grain fill duration, plant height and grain yield. Selection for early flowering/maturity in early segregating generations would be expected to result in genetic improvement towards earliness in high latitude spring wheats. Incorporation of the vernalization responsive gene Vrn-B1 in combination with vernalization non-responsive gene Vrn-A1 into spring wheats would aid in the development of early maturing cultivars with high grain yield potential for the high latitude wheat growing regions of the northern hemisphere.     

Seed formation in triploid loquat (Eriobotrya japonica) through cross-hybridization with pollen of diploid cultivars

As the fruits of loquat (Eriobotrya japonica, 2n = 2x = 34) carry large seeds, the breeding of seedless loquat has long been a goal. The recent creation of triploid cultivars (2n = 3x = 51) and the application of gibberellins allow commercial production of seedless loquat, but the possibility of seed formation in triploid loquats has not been carefully investigated. Through crossing experiments and cytological observations of meiosis and pollen tube growth, we found that the triploid line 3N-N28 was essentially self-sterile, but developed seeds on pollination with pollen from diploid cultivars at rates of up to 5.5%. Almost half of the seedlings survived to 5 months, and carried diploid (2n = 34), tetraploid (2n = 68), or aneuploid chromosome numbers. Our results suggest that triploid loquat cultivars might retain the risk of seed formation. Protection from pollination by diploid cultivars or the development of new triploid cultivars will be necessary to ensure the production of seedless loquat fruits.     

木质素生物合成及其基因调控的研究进展

木质素作为陆地上含量丰富的高分子有机物之一,对植物的结构与防御功能都具有重要作用。然而,它的存在也给制浆造纸工业,苎麻纺织工业及畜牧业生产带来负面影响。因此,对于木质素的生物合成途径及其基因调控的探索已成为研究热点。木质素的生物合成途径十分复杂,存在多基因、多途径的交互作用。利用基因工程技术调控木质素生物合成途径中的关键酶基因的表达可以降低木质素含量或者改变木质素组成成分,从而开发新型植物资源,从源头降低成本,减少污染。本文介绍了修订后的木质素生物合成途径,重点阐述其基因调控的研究成果。并针对应用到实际生产中存在的问题,在木质素调控基因的选择、多基因遗传操作、启动子的选择、转基因技术的选择等方面提出一些可行的建议。     

两种苎麻纤维素合酶基因cDNA序列的克隆及表达

从苎麻转录组数据出发, 利用Blast工具从中分析出与多种植物纤维素合酶高度相似的片段CL789和Unigene20360。根据片段信息设计特异性引物, 从苎麻[Boehmeria nivea (Linn.)Gaud.]栽培种湘苎3号中克隆纤维素合酶核心片段, 并利用5'及3'RACE技术获得2个片段的全长cDNA。两者都具有典型的纤维素合酶特征结构域, 表明为2个苎麻纤维素合酶基因CesA的cDNA序列, 分别命名为BnCesA2和BnCesA3。BnCesA2基因编码区全长度3240 bp, 编码1 079氨基酸多肽; BnCesA3基因编码区全长3120 bp, 编码1039氨基酸多肽。对BnCesA2和BnCesA3基因在湘苎1号、湘苎3号、湘潭大叶白和城步青麻苎麻品种木质部和韧皮部荧光定量PCR分析显示, 2个基因在不同品种苎麻的木质部及韧皮部都有表达, 但表达量存在着一定差异, 整体而言BnCesA2具有更高的表达水平, 其木质部和韧皮部的表达都为BnCesA3的2~5倍。推测BnCesA2和BnCesA3都参与了苎麻细胞壁的次生合成。     

基因工程在葡萄研究中的应用进展

简述了葡萄常规育种方法存在的问题,回顾了近几年国内外通过基因工程技术在葡萄转基因体系的建立和基因功能研究2个方面取得的最新研究进展,分析了目前葡萄基因工程研究中的主要问题,并对葡萄基因工程的前景进行了展望。认为基因工程的研究方法为葡萄抗病抗虫、高产优质等优良品种的选育开辟了广阔的途径,葡萄育种学家以此为契机,将会培育出更多的优良品种。     

Improving in vitro induction of autopolyploidy in grapevine seedless cultivars

The efficiency of in vitro polyploidization depends on several variables associated to the plant, the antimicrotubule agent and the interactions between them. In the present work, we have used response-surface methodology to determine the best operating conditions for plant recovery in polyploidization assays for shoot apices and somatic embryos of two seedless grape cultivars, employing colchicine and oryzalin. Explant type, tubulin-interfering compound and concentration were the critical factors determining plant recovery. Linear reduction in viable plant regeneration via organogenesis and somatic embryogenesis was obtained by increasing oryzalin concentrations and treatment time, whereas the effects of colchicine were better described by a quadratic design for both explants types. The conditions promoting higher rates of plant recovery were used in chromosome doubling experiments with oryzalin and colchicine for shoot apices and somatic embryos of ‘Crimson seedless’ and ‘BRS Clara’. The established protocols allowed the recovery of non-chimerical autotetraploid plants at rates higher than 30 % for both cultivars. Stomata size parameters statistically correlate to the ploidy level of the regenerants and were effective for preliminary polyploidy screening. Autotetraploid lines of seedless grapes were incorporated into the Vitis germplasm bank for further agronomical evaluations. To our knowledge, this is the first report of in vitro oryzalin induced polyploidization of grapevine and of the use of mathematical modeling to optimize chromosome doubling in plants.     

胚挽救技术创造无核抗病葡萄新种质

为提高无核抗病、优质葡萄新种质的选育效率,以无核欧洲葡萄品种作母本,中国野生刺葡萄‘塘尾’、‘雪峰’、山葡萄‘双优’及欧山杂交品种‘北醇’做父本,利用胚挽救技术对取样时间和培养基进行优化;用分子标记筛选后代的无核与抗病性状。结果表明：（1）15个杂交组合中共获得杂交果实13341个,接种幼胚珠8589枚,幼胚发育胚1338枚,平均胚发育率15.59%,正常成苗234株,平均成苗率2.73%,‘红宝石无核’ב塘尾’组合胚挽救效果最好,胚发育率为40.17%、成苗率7.76%和成苗植株数178株。（2）‘火焰无核’×北醇’授粉后42天取样,胚发育率(11.37%)和萌发率(3.52%)为最高。‘京可晶’ב北醇’授粉后38天取样,胚发育率(4.3%)和萌发率(1.08%)为最高。（3）以MM3为基础培养基,外源添加3 mmol/L的腐胺,‘红宝石无核’ב塘尾’获得胚发育率(55.83%)和成苗率(20%)为最高。（4）4个自然授粉的品种（‘昆香无核’、‘美丽无核’、‘红宝石无核’、‘赫什无核’）,其胚珠在E20A培养基中更有利于幼胚发育率提升。（5）使用葡萄无核标记SCF27-2000和GSLP1-569、抗霜霉病标记S294-369和S382-615及抗白粉病标记OPY13-661进行筛选,杂种后代中有16株同时携带无核且抗霜霉病与抗白粉病特异性条带。该试验为无核葡萄抗病育种提供了新材料,将为进一步开展中国野生葡萄育种相关研究奠定基础。     

Germplasm Variability and Environmental Effects on Stem Cellulose and Lignin Concentrations in Alfalfa*

Stem cellulose and lignin concentrations are major determining factors of alfalfa (Medicago sativa ssp. sativa L.) forage quality. Only limited information is available on the genetic variability and the influence of environmental effects on these two stem-quality traits. Our objectives were to: evaluate the variability for stem cellulose and lignin concentrations in modern alfalfa germplasms across several harvests; observe the environmental stability of these two quality traits in 32 alfalfa clones selected high or low for either stem cellulose or lignin concentration; and examine the relationships between these two stem-quality traits and leaf and stem crude protein (CP). Fifty alfalfa entries (cultivars and experimental populations) were established May 1993, and sampled for stem acid-detergent lignin (ADL), stem acid-detergent cellulose (ADC), and stem CP on Sep 1993, June and Aug 1994. Clones were vegetatively propagated from individual plants selected for extremes in stem ADL and ADC and transplanted into blocks at two locations in May 1994 and sampled for quality analysis in September 1994. For all samples, leaves were hand-separated from stems and stem ADL, ADC, CP and leaf CP concentration were determined using near infrared reflectance spectroscopy. Entry differences for stem ADL and ADC were detected only at the June 1994 harvest date in the cultivar study. Spearman's ranked correlations over years in the clonal study demonstrated greater environmental stability for stem ADC (r = 0.70, P ≥ 0.01) than for stem ADL (r = 0.54, P # 0.05). Environmental effects had an impact on both traits, but stem ADC showed greater potential for improving forage quality. Simple correlations showed that decreasing stem ADL or ADC would have minimal effect on leaf CP and may increase stem CP.     

Flavan-3-ols and procyanidins in grape seeds: biodiversity and relationships among wild and cultivated vines

Flavan-3-ol monomers and polymers composition of seeds from wild (17) and autochthonous (8) Vitis vinifera grapes growing in northern Tunisia were evaluated. Wild grape seeds were spherical with a small beak and relatively a high seed/berry ratio (~ 18.1%w/w). Local cultivars developed pyriform-shaped seeds with a well-developed beak representing on average 2.2% of total weight of the berry. Flavanol concentrations ranged between 40.9 and 67.5 mg/g FW in seeds from wild accessions and between 48.9 and 96.7 mg/g FW in seeds from cultivated grapes. Differences between accessions were highly significant (p < 0.01) and seeds from cultivar ‘Boukhasla’ showed the highest polyphenols content. Among flavan-3-ol monomers, (+)-catechin was predominant for all ecotypes and generally their abundance was: (+)-catechin (Cat) > (?)-epicatechin (Ec) > (?)-epicatechin-3-O-gallate (EcG). The Cat/Ec ratio was approximately 1.7 for wild grapes while it was about 2.5 for cultivated grapes. Procyanidins in wild seeds differed from cultivated ones by a lower mDP and higher proportions of galloylated derivatives, likely to affect fruit bitterness and astringency. (?)-epicatechin was the main extension subunit in grape seed procyanidins, reaching on average 52% in wild and 58% in cultivated seeds. Hierarchical cluster analysis based on seeds morphometry and procyanidin profile indicated close proximity between some wild and cultivated grapes suggesting that some cultivars derived from ancestral events of local domestication or cross hybridization with native wild plants.     

菜用大豆品种花荚粒形成的初步研究

通过对24个菜用大豆品种花荚及籽粒形成的比较,认为不同熟期类型品种间发育规律存在差异。早熟品种较晚熟品种花、荚形成发育快,时间短,脱落也快。籽粒形成主要集中在三个时期：籽粒重在中后期,荚皮重在中前期,荚重在中期。中期的生殖生长是籽粒形成的基础,也是影响产量的关键。因此在江苏徐淮地区的生态条件下,鼓粒前的花荚期长短是选择菜用大豆品种生育指标的重要因素。菜用大豆鼓粒前的花荚期以20d左右为宜。     

枯萎病菌诱导感、抗陆地棉品种的基因表达谱分析

选用陆地棉抗枯萎病品种中棉所12和感枯萎病品种新陆早7号,利用Solexa高通量测序技术分析棉花幼苗受到枯萎病菌侵染后3 h和6 h根部组织的基因表达谱。中棉所12受病菌侵染后3 h与0 h相比,6 h与0 h相比以及6 h与3 h相比,分别鉴定出4447、5481和2559个差异表达基因;新陆早7号分别鉴定出8615、6727和2078个差异表达基因;2个品种比较,在3 h和6 h分别鉴定出1879个和500个差异表达基因。基因功能分析表明,差异表达基因划分为生物学过程、细胞组分和分子功能注释本体,并进一步细分为48个功能类别。代谢通路分析显示,中棉所12和新陆早7号在枯萎病菌侵染后的6 h与0 h相比鉴定出的代谢通路(Pathway)最多,均有126条;在枯萎病菌侵染后6 h,2个品种相比鉴定出的代谢通路最少,仅有89条。各个比对组的代谢通路涉及的基因可被划分为次生代谢产物的生物合成、多糖的生物合成和代谢、环境适应和免疫系统等13类。在环境适应和免疫系统分类中存在着植物与病原菌相互作用代谢通路,涉及到996个已知功能的差异表达基因,有444个基因上调表达,552个基因下调表达。其中,差异表达基因最多的是WRKY转录因子家族基因,其次为丝氨酸/苏氨酸蛋白激酶基因,而DNA损伤修复/耐受性蛋白,JAZ1、RAR1和RPM1互作蛋白,S位点的特异性糖蛋白S6前体以及钙牵引蛋白等6类基因参与该代谢通路的数目最少。最后随机抽取6个基因进行实时荧光定量PCR验证,其结果与测序结果一致。     

Current status and future scenarios of hemp breeding

Summary Hemp is a multi-use crop, able to provide fiber, cellulose, seeds and seed oil, cannabinoid, and biomass. Integrating many agroindustrial chains, Cannabis is considered a crop model in which insights into specific metabolic pathways and biosynthetic processes are valuable for improvement of the plant for all sets of industrial derivatives. In this review the hemp breeding status is elucidated and many aspects are focused: (i) recovering, maintenance and characterization of genetic resources; (ii) widening of germplasm and genetic variability; (iii) marker-assisted selection and development of breeding programs; (iv) sexual differentiation; (v) monitoring of THC content. Modern hemp varieties for fiber and other specific end uses have been developed and new varieties are entering production. The scenario for the breeding advances in hemp relies on basic and applied research which provides insights to identify a strategy for the design of modified plants with enhanced performance. This is accounted by the dissection of traits into components and the modification of single steps of the related metabolic pathways. These advances are provided by genomic techniques and are able: (i) to identify key genes encoding enzymes and regulatory factors participating in cannabinoid, fiber and oil biosynthesis; (ii) to identify the mode of regulation of these genes; (iii) to characterize the function of the selected genes through higher, lower or specific expression incited by specific promoters. The identification of molecular markers for specific traits, gathered in a saturated linkage map, will have a remarkable impact on hemp breeding. The advances in basic and applied research make it possible to design methods for the identification of superior parents and cross combinations and the development of selection schemes that rely on less labour-intensive and time-consuming methods.     

木质素生物合成途径中关键酶基因的分子特征

本文主要对苯丙氨酸裂解酶（phenylalanine ammonia-lyase,PAL）基因、4-香豆酸辅酶A连接酶（4-coumarate-CoA ligase,4CL）基因、肉桂醇脱氢酶（cinnamyl alcohol dehydrogenase,CAD）基因、过氧化物酶（peroxidase,POX）基因、漆酶（laccase,LAC）基因、dirigent（DIR）蛋白基因等木质素生物合成途径中关键酶基因的克隆、表达、调控等研究进展进行综述,旨在揭示上述木质素生物合成途径中关键酶基因的分子特征,为通过转基因技术来调控植物体中木质素的含量及其化学组成从而得到改良的新植物资源提供思路。     

Rachis browning in four table grape cultivars as affected by growth regulators or packaging

Plant growth regulators (PGRs) are used to increase berry size in table grapes. The objective in this study was to determine if the levels of PGRs used commercially affected rachis postharvest quality. Three seedless table grape cultivars, ‘Mystery’, ‘Superior’, and ‘Crimson’ were treated with gibberellin (GA), cytokinin (CPPU), or both at berry diameter of 6–8 mm, while a seeded cultivar, ‘Redglobe’ was treated at 13 mm. The fruits were harvested at commercial maturity and held for 7 days shelf-life either immediately after harvest or after 2 weeks storage at 0 °C. The combination of GA and CPPU increased berry weight and diameter, and rachis diameter in all three seedless cultivars, but had minor or no effect on TSS and acidity. Rachis quality was measured by a browning scale derived from image analysis. Browning was lower after storage for rachis of ‘Mystery’ treated with GA, while the other three cultivars showed no difference between PGR treated and control rachis. Reduction of water loss by covering punnets of control clusters with microperforated film had a significant positive effect on decreasing weight loss in all cases and a positive effect on slowing rachis browning. Calculation of the shelf-life required to reach 50% rachis browning found that shelf-life was longer for the microperforated packaging as compared to open packages in all the cultivars, with stronger influence during shelf-life after storage than shelf-life after harvest. However, the data also indicated that rachis browning cannot only be attributed to weight loss, either from the whole cluster or from the rachis.     

中国东北大豆与日本大豆品种PEPCase基因的多态性

Soybeans in Northeast of China are one of the most importantgenetic resources in the world.In order to use themeffectively insoybean breeding,it is necessary to evaluate the maincharacteristics,especially the quality characteristics such as seedprotein content.It is reported that the PEPCase activity in soybeanseeds is positively correlated with the seed protein content andnegatively correlated with the lipid content[1].PEPCase is alsopresent in seeds of different species[2~4],and may play…     

Agronomical and quality traits of runner bean germplasm and implications for breeding

White runner bean landraces are greatly appreciated in the North highlands of Spain due to their excellent culinary seed quality. Runner bean cultivars are grown like pole beans. Diversity within a runner bean collection of 31 accessions from the Iberian Peninsula (Spain and Portugal) was examined using morphological, agronomical and seed quality traits. Landraces showed significant differences for most of the agronomical and seed quality traits studied except for seeds per pod, water absorption, seed coat tenderness and floury texture. Runner bean landraces showed sufficient variability to select inbred lines for future breeding. Genotype × environment interaction was significant for days to first flowering, days to first dry pod, seeds per pod and seed length. The majority of physical and nutritional seed quality traits studied which are important to determine the commercial value of a variety were not subject to environmental influences. Different selection pressures affecting to the runner bean genetic material could have occurred in several regions of the Iberian Peninsula. Extra-large and high yielding runner bean germplasm was identified and represents a valuable source of genetic diversity that has potential for development of improved cultivars to be chosen for commercialisation. This revised version was published online in July 2006 with corrections to the Cover Date.