16份石榴RAPD扩增产物的两种电泳方法检测及其序列特征

本实验以16个石榴品种为实验材料,筛选出10个重复性及多态性均较好的引物进行RAPD分析。分别采用琼脂糖凝胶以及聚丙烯酰胺凝胶(PAGE)电泳检测方法对PCR扩增结果进行检测并对其结果进行比较,结果显示,两种电泳方式均能得到较为清晰的扩增条带,且两种电泳方式获得的条带总数及多态性条带数均有所不同,琼脂糖凝胶电泳方法共检测出76条带,其中有43条为多态性谱带,多态性比率为56.4%;而在PAGE电泳方法共检测出123条谱带,多态性谱带数为87条,多态性比率为70.95%。PAGE电泳方法检测出的条带数约为琼脂糖凝胶电泳方法检测出条带数的1.5倍。基于两种电泳方法所得RAPD标记的多态性位点,利用NYSYS软件计算遗传相似系数,并构建遗传关系聚类图,分析结果显示,石榴遗传多样性丰富,两种电泳方法所得聚类结果大致相同,可以利用RAPD分子标记及两种电泳检测方法对不同数量的石榴进行分子水平的品种鉴定和遗传多样性的分析。同时通过对来自几个引物随机挑选的17个片段进行克隆,测序结果显示17个片段都是对应引物的RAPD扩增产物,其中有3条是编码蛋白的基因片段,表明了RAPD不仅扩增基因组上的非编码蛋白序列,同时也可以扩增编...     

普通小麦(T.aestivum L.)Waxy蛋白种质资源研究

用SDS-PAGE方法对国内外293份小麦品种(系)Waxy蛋白进行了鉴定，筛选出缺Wx-A1的材料2份；缺Wx-B1的材料15份，其中14份材料为本研究首次发现；缺Wx-D1的材料2份；同时缺失Wx-A1和Wx-B1的材料2份；全缺的材料3份。利用分子标记对不同的Waxy蛋白缺失类型进行了鉴定，开发了一个Wx-A1位点的共显性STS标记。     

我国棉花大丽轮枝菌(Verticillium dahliae)电泳图谱分析

对24株来自不同地区、作物的不同致病类型的大丽轮枝菌,进行了同工酶和可溶性蛋白电泳图谱分析。结果表明,24株大丽轮枝菌苹果酸脱氢酶酶谱具有种的特异性,酯酶酶谱具有属的特异性。谷氨酸脱氢酶、过氧化物酶、超氧化物歧化酶和可溶性蛋白电泳图谱与病菌的致病类型有关,可将24株大丽轮枝菌分为2群,群Ⅰ包括20个菌株,均属于中等致病力或弱致病力的非落叶型菌系;群Ⅱ包括4株菌,均属于强致病力的落叶型菌系。     

小麦高分子量谷蛋白亚基与小麦品质性状关系的研究

采用SDS-PAGE方法, 通过对4个优质亲本与3个农艺亲本间杂交获得的F2群体的每一单株及其亲本进行小麦高分子量谷蛋白亚基(HMW-GS)组成分析, 并对每一F2单株上F3籽粒群体的高分子量谷蛋白亚基组成与其籽粒蛋白质含量、 SDS-沉降值的关系进行研究,结果表明：小麦高分子量谷蛋白亚基组成不同的群体间籽粒蛋白质含量差异不显著     

Modification of physicochemical properties and in vitro digestibility of wheat flour through superheated steam processing

Native and moistened wheat flours (moisture contents were 13.5 and 27.0%, respectively) were treated with superheated steam (SS) at different temperatures (140 and 170 °C) and times (1, 2 and 4 min). Their physicochemical and digestive properties were analyzed. For native flour, SS treatment altered the starch molecular structure and behavior slightly. While for moistened flour, crystalline degree, gelatinization enthalpy, amylose leaching (AML) and falling number significantly decreased, but thermal transition temperatures increased with the rise of treating severity. Clumping of starch granules, aggregation of proteins and formation of amylose-lipid complexes occurred in both native and moistened flours. Broader pasting temperature ranges and higher viscosities were found on SS-modified flours. Additionally, SS treatment on moistened flours increased resistant and slowly digestible starch contents. In general, SS treatment induced changes in starch molecular structure and reactions among flour components leading to more stable structures, thus affecting their pasting behavior, thermal properties and in vitro digestion.     

The impact of redox agents on further dough development,relaxation and elastic recoil during lamination and fermentation of multi-layered pastry dough

The role of gluten proteins during lamination and fermentation of multi-layered wheat flour pastry dough was examined by including oxidizing or reducing agents in the recipe to respectively strengthen or weaken the gluten protein network. Pastry burst rig textural measurements showed that dough strength increases during lamination up to 16 fat layers. However, further lamination up to 64 and 128 fat layers decreases the dough strength, most likely due to destruction of layer integrity. Redox agents strongly affect dough strength. Furthermore, fermentation and spread tests showed that they strongly influence elastic recoil immediately after lamination and during relaxation. Moreover, elastic recoil consistently occurs to a greater extent in the final direction of sheeting. None of the observed changes in dough strength and relaxation behaviour could be linked to changes in the levels of protein extractable in sodium dodecyl sulfate containing medium (SDS-EP). This suggests that changes occur preferentially either within the SDS-extractable or within the non-SDS-EP fraction and that they do not render non-extractable protein fractions extractable or vice versa. Furthermore, elastic recoil is most likely caused by reformation of inter- and intramolecular hydrogen bonds and hydrophobic interactions.     

Antimicrobial activity of partially characterized analytes from Bacillus pumilus(B2)

This study was carried out to characterize the antimicrobial substance produced by the strain of Bacillus pumilus (B2) obtained from Novozymes Biologicals Inc. to compare its antimicrobial activity by agar well diffusion assay and bacteriocin activity assay via critical dilution method against seven different strains of Vibrio spp., specifically V. alginolyticus (A01), V. cholerae (C01), V. fluvialis (F01, F02), V herveyii (H), V. mimicus (M01), V. parahaemolyticus (P01) and V. vulnificus (V01, V02). All Vibrio spp. were isolated from the hemolymph and intestine of the white faeces disease‐infected moribund pacific white‐leg shrimp Litopenaeus vannamei (Boone 1931) and one strain (V. harveyi) from its diseased postlarva. The cell‐free neutralized supernatant (CFNS) of B2 showed moderate thermo‐stability being stable up to 70°C for 60 min with, however, reducing activity above 80°C for 20 min. B2 antimicrobials showed a stable activity within the pH ranging from 6 to 10 at room temperature and at 4°C, while residual antimicrobial activity of crude CFNS showed tolerance to salinity up to 7% of sodium chloride below 4°C. No B2 activity was obtained while exposed to proteolytic enzyme, such as proteinase k and pepsin, while its activity kept stable exposed to lipase. Initial B2 characterization for antimicrobial substance in CFNS revealed proteinaceous in nature owing to activity loss against proteolytic enzymes and no lipid moiety activity against lipase, which could be categorized as bacteriocin‐like inhibitory substance having potential application against several strains of Vibrio spp. in aquaculture.     

微卫星产物变性与非变性PAGE-银染方法比较

用哲罗鱼的微卫星引物对哲罗鱼的2个微卫星位点进行PCR扩增。将扩增产物在变性与非变性聚丙烯酰胺凝胶(PAGE)上电泳,银染后发现微卫星扩增产物在二者上的电泳结果存在明显差异。在非变性凝胶中存在较多非特异条带,而在变性凝胶中微卫星扩增产物条带清晰,易于鉴定。     

Thermostability variation in alleles of barley beta-amylase

Thermostability assays in conjunction with IEF and molecular mapping were used to identify three beta-amylase alleles (Bmyl-Sd1, -Sd2L, -Sd2H) in cultivated barley and an additional allele (Bmy1-Sd3) in an accession of wild barley Hordeum vulgare ssp. spontaneum. The four forms of beta-amylase exhibit different rates of thermal inactivation in barley extracts. This variation was shown to persist after the proteolytic processing of the enzyme that occurs during germination. Three forms of beta-amylase representing the range of thermostabilities were purified and shown to have T₅₀ temperatures of 56·8°C for the Sd2L enzyme, 58·5°C for the Sd1 enzyme, and 60·8°C for the Sd3 beta-amylase from wild barley. Analysis of the relationship between beta-amylase thermostability and fermentability, i.e. the yield of fermentable sugars obtained from starch hydrolysis during brewing in 42 commercial malt samples suggests that increased thermostability results in more efficient starch degradation. Screening for specific beta-amylase alleles is proposed as a method for increasing fermentability in malting barley.     

Genetic analysis of late blight resistance in a new RIL population of tomato derived from LB-resistant Solanum pimpinellifolium accession PI 270443

Late blight (LB), caused by Phytophthora infestans, is one of the most devastating diseases of tomato (Solanum lycopersicum) worldwide. Aggressive pathogen isolates resistant to fungicides have driven research in favour of finding new sources of host resistance for tomato breeding. Recently, we reported S. pimpinellifolium accession PI 270443 exhibiting LB resistance stronger than all commercial LB-resistant tomato cultivars. The purpose of this study was to examine the inheritance of LB resistance conferred by this accession. An interspecific cross was made between PI 270443 and a LB-susceptible tomato breeding line and advanced to F₁₀ generation. A total of 166 F₉ and corresponding F₁₀ recombinant inbred lines (RILs) were evaluated for response to LB in four replicated greenhouse experiments. Estimates of heritability (h²) of LB resistance, determined by parent–offspring (F₉:F₁₀) correlation analysis, ranged from 0.66 to 0.81, with an average of 0.76. The moderately high h² of LB resistance in PI 270443 suggests the utility of this accession for tomato breeding. Molecular mapping and RNA-sequencing efforts are underway to identify genes underlying LB resistance in PI 270443.