甜菜DAMD引物的筛选及不同凝胶系统对扩增产物多态性的影响

为了筛选适宜于甜菜品种纯度鉴定的DAMD引物,笔者利用12 个不同的甜菜品种分别对26 条DAMD引物进行扩增,并分别采用6%聚丙烯酰胺凝胶电泳和2%的琼脂糖凝胶电泳对扩增产物进行检测。结果表明,从26 条DAMD引物中筛选出16 条扩增清晰的引物,这16 条引物共扩增出139 条带,其中多态性条带为122 条,多态性百分比为87.7%,这16 条引物可以作为甜菜品种纯度和真实性鉴定的核心引物,同时发现聚丙烯酰胺凝胶电泳与琼脂糖凝胶电泳的检测结果差异不大,而琼脂糖凝胶具有制作方便、检测简单以及不使用任何有毒药剂等优点,推荐甜菜DAMD扩增产物的检测使用琼脂糖凝胶电泳。     

Introgression of antioxidant activity into cassava (Manihot esculenta C.): an effective technique for extending fresh storage roots shelf life

Physiological postharvest deterioration (PPD) of cassava is the main constraint affecting its nutritional and economical values. PPD is induced by wounds when detaching storage roots from mother plant during harvesting. It is accelerated by the reactive oxygen species (ROS) such as oxygen ion (O₂) and peroxide (O₂)^?2. The carotenoid content and its antioxidant property can help in extending shelf life of cassava storage roots. The primary mode of action of carotenoids as antioxidant is to quench singlet oxygen. Cassava breeding was reported to successfully introgress and improve carotenoid content into cassava. The two types of phytoene synthase (PSY) enzymes (PSY1 and PSY2) are key regulators of carotenoids accumulation in cassava. Carotenoids formation and accumulation in cassava storage roots are induced by a single nucleotide polymorphism in PSY2 which causes a non‐conservative amino acid exchange. This single nucleotide polymorphism in PSY gene is co‐segregated with β‐carotene in cassava storage roots, a phenomenon that could help to unravel the mechanism of introgression of carotenoids into cassava. This article investigates breeding feasibility for improving quality of cassava landraces in developing countries.     

甘蓝型油菜抗裂角品种(系)的筛选与分析

利用随机碰撞测试方法, 对229份自交纯合的甘蓝型油菜品种(系)进行了抗裂角指数测定, 以期筛选出抗裂角种质。结果表明, 抗裂角性状在现有品种(系)中存在广泛变异, 变异系数达114.4%。发现了2份抗裂角的品种(系), 占0.9%; 较抗的资源占3.93%; 处于中间状态的品种(系)占8.73%; 易裂角资源占27.07%, 极易裂角的品种(系)占59.39%。选择6个品种(系)进行了连续3年的测试, 表明抗裂角性状由品种(系)的遗传特性决定, 但受环境条件影响; 随机碰撞法具有较好的重现性。简单相关分析显示, 抗裂角指数与角果密度呈显著负相关, 与角皮厚度、角果长度、角果宽度、角喙长度、角粒数呈显著正相关, 但相关系数都很小。     

荞麦轮纹病抗性鉴定方法的建立及荞麦抗病种质资源的筛选

为建立荞麦轮纹病的抗性鉴定方法和筛选可利用的抗轮纹病的荞麦种质资源,本研究以TP2和KP14为供试材料,在健康荞麦叶片正面针刺处理,然后用荞麦轮纹病的病原菌菌饼进行接种;利用建立的抗性鉴定方法,对50份荞麦种质资源进行抗病性评价。结果显示:针刺5针接种叶片病斑出现较快,且比针刺1针、针刺3针的叶片病斑大,针刺5针的病情指数最高。50份荞麦种质资源的病情指数有差异,其中YZ-18的病情指数为29.63,为高抗种质资源,YZ-2、YZ-5、YZ-13和YZ-9的病情指数均低于40,为中抗种质资源,其余为感病种质资源。因此,接种前叶片正面针刺5针的方法为最佳的抗性鉴定方法。50份抗性评价的荞麦种质资源中,筛选出5份抗病种质资源。     

Instrumental and sensory quality characteristics of blueberry fruit from twelve cultivars

We compared the instrumental and sensory quality characteristics of blueberry fruit from ten highbush (Vaccinium corymbosum L.) cultivars, Chanticleer, Weymouth, Hannah's Choice, Duke, Bluecrop, Coville, Berkeley, Bluegold, Elliott and Lateblue and two rabbiteye (Vaccinium virgatum Aiton) cultivars, Coastal and Montgomery, grown in New Jersey. Cultivars varied in sensory intensity and acceptability scores. Highbush cultivars, Coville and Hannah's Choice, scored highest among the cultivars in sensory scores for intensity of blue color, juiciness, sweetness and blueberry-like flavor and for acceptability of appearance, color, fruit size, sweet/tart balance, flavor and overall eating quality. In contrast, rabbiteye cultivars, Coastal and Montgomery, and the highbush cultivars, Elliott and Weymouth, scored lowest among the cultivars in sensory scores for intensity of bursting energy, skin toughness, texture during chewing, juiciness, and blueberry-like flavor and for acceptability of appearance, color, fruit size, flavor and overall eating quality. Analytical quality characteristics of surface color, size, compression firmness, soluble solids content (SSC), pH, titratable acidity (TA), SSC/TA ratio, and aromatic volatile concentration also varied among cultivars, but no instrumental measurement adequately predicted consumer acceptability scores. The overall eating quality of blueberry fruit was best correlated with flavor scores followed by sensory scores for intensity of juiciness, bursting energy and sweetness and for acceptability of appearance.     

Recurrent breeding method enhances the level of blackspot (Didymella pinodes (Berk. & Blox.) Vestergr.) resistance in field pea (Pisum sativum L.) in southern Australia

Blackspot, caused by Didymella pinodes (Berk. & Blox.)Vestergr., is one of the most important diseases of field pea, causing significant reduction in seed yield and quality in southern Australia and in other parts of the world. Development of resistant germplasm has been slow because of the low level of resistance found in the available germplasm, poor reliability of screening methods and the polygenic nature of inheritance. Crosses were made between agronomically suitable lines and resistant germplasm from different sources. Their progeny were advanced through the single seed descent method and single plants were selected at F₄/F₅. The F₄/F₅ derived lines were screened against blackspot in the field under disease pressure and evaluated for grain yield at multilocations over 2 years. Despite the low level of resistance in the parental germplasm, the level of resistance has increased significantly in the new germplasm. Many of the resistant lines were late and low yielding, but lines with higher resistance and early flowering and high yield potential were also identified indicating that the disease resistance, adaptation and yield potential can be combined. However, the resistance identified in this study is only partial and suitable agronomic practices may need to be supplemented to minimise the yield loss and enhance the benefits of this partial resistance.     

普通小麦品种小偃54中α/β-醇溶蛋白编码基因的克隆与序列分析

醇溶蛋白是小麦籽粒贮藏蛋白的重要组分,其组成与含量对小麦加工品质具有重要影响。本研究建立了利用PCR从普通小麦基因组BAC文库中筛选含有α/β-醇溶蛋白基因序列BAC克隆的方法,并获得9个不同的含有α/β-醇溶蛋白基因的BAC克隆。从其中鉴定出17个α/β-醇溶蛋白基因,其编码区序列长度为852~957 bp。12个序列在编码区内存在提前终止密码子,推测为假基因。其他5个成员(Gli-Xy54-1、Gli-Xy54-2、Gli-Xy54-3、Gli-Xy54-7和Gli-Xy54-13)分别编码291、310、311、287和317个氨基酸残基,都具有α/β-醇溶蛋白一级结构的典型特征。根据推导的氨基酸序列中乳糜泻病诱发因子的分布情况及多聚谷氨酰胺重复区的长度差异,推测Gli-Xy54-7可能定位于6A染色体,Gli-Xy54-2、Gli-Xy54-3和Gli-Xy54-13可能定位于6B染色体,Gli-Xy54-1可能定位于6D染色体。基因聚类分析支持了上述推论。这是第一次从普通小麦中筛选到包含α/β-醇溶蛋白基因的BAC克隆,并从中得到目标基因全长,对进一步研究普通小麦基因组中α/β-醇溶蛋白编码基因的组成、表达与功能有较好的参考价值。     

内生放线菌Fq24的拮抗筛选及其代谢产物生物活性研究

通过对番茄内生放线菌的拮抗筛选和其代谢产物生物活性的研究,为防治番茄灰霉病寻找新的生防菌源,并探索植物内生放线菌的生防机理。从健康番茄植株上分离到9株放线菌。通过平板对峙培养和发酵液抑菌活性测定、筛选对番茄灰霉菌有拮抗作用的菌株。以番茄种子及幼苗生长情况,防御酶活性变化,盆栽试验对其代谢产物的促生作用、抗病诱导作用、防病作用等生物活性进行了研究。结果表明：（1）菌株Fq24对番茄灰霉菌的抑菌圈半径大于10 mm,其发酵液的抑制作用可达到68.4%。（2）中浓度的Fq24发酵液促进根茎的生长,幼苗的干、鲜重及株高最大为对照的168.35%、286.30%和70.79%,均为显著增加。（3）番茄幼苗体内防御酶活性明显提高,POD、SOD活性最大增加为71.3%和62.3%。（4）喷施Fq24发酵液可明显降低灰霉病的发病率,取得82.14%的防效。Fq24是株有拮抗作用的菌株,其代谢产物能显著促进番茄种子和幼苗生长;诱导番茄体内抗性酶活性提高,增加植物抗性;明显降低发病率,防病效果好。     

Barley mutants with increased tolerance to aluminium toxicity

Acid soil and associated aluminium toxicity are considered as the number one abiotic factor limiting crop production. Over 2 billion hectares of acid soils exist world-wide, both in tropical and moderate climatic zones. In Poland acid soils represent up to60% of arable land. At soil pH < 5.0 Al ions become soluble in water and toxic to plants. Genetic improvement of Al tolerance in crops is the only alternative to soil liming, a traditional but short term and expensive agricultural cure to raise soil pH. Of the various cereals, barley is the most sensitive to Al toxicity. The known sources of Al tolerance in barley are limited to old cultivars and landraces. While they represent multiple alleles of a single locus, there is no potential to improve Al tolerance through recombination of non-allelic additive genes. In the Department of Genetics, Silesian University we have employed induced mutations for rapid creation of variability for Al tolerance in barley. Thirteen mutants with increased levels of tolerance to Al toxicity have been selected in M₃ generation after mutagenic treatment of four barley varieties with N-methyl-N-nitroso urea (MNH) and sodium azide. Six further Al tolerant mutants were identified in the collection of semi-dwarf mutants of the Department. All selected mutants confirmed Al tolerance with the use of three different methods of screening, i.e., root re-growth, root tolerance index and hematoxylin staining. Fourteen mutants exhibited significant root re-growth after 48 hour incubation with 3 ppm Al⁺³ and two of them, namely RL819/2 and RL820/6 were tolerant even to 6 ppm Al⁺³. Crosses of two selected mutants with their respective parent varieties indicated that Al tolerance in each mutant was controlled by a single recessive gene. Out of three methods tested, the root re-growth method facilitated by hematoxylin staining proved to be the most reliable technique for large scale testing. Double treatment with MNH or combined treatment with sodium azide and MNH and 6hinter-incubation germination between treatments were the most successful treatment combinations for induction of aluminium tolerance in barley. This revised version was published online in August 2006 with corrections to the Cover Date.     

双丙氨磷在橡胶树遗传转化中的应用

本研究以GV3101::pMP90RK为工程菌株,bar基因作为筛选标记基因,研究了双丙氨磷在橡胶树根癌农杆菌介导法转基因体系中对转基因愈伤组织的筛选效果,并利用该体系将橡胶树转录因子HbWRKY5转入橡胶树无性系热研8-79中。实验结果表明,3mg/L双丙氨磷可作为橡胶树转基因体系中抗性愈伤组织筛选的最佳浓度。本研究中共获得了3个抗性愈伤组织系,抗性愈伤经过体细胞胚发生共获得234个体细胞胚,经植株再生培养后共获得22株再生植株。对再生植株进行的PCR鉴定结果表明,所获得的再生植株均为转基因植株。因此,bar基因结合双丙氨磷的筛选体系可以作为橡胶树转化体系中的有效筛选体系。