番茄成熟突变体(rin,nor,alc)及其F_1的贮藏生理特性研究

本研究对番茄成熟突变体rin、nor、alc材料与正常成熟番茄品种及其杂种一代果实,在绿熟期采收后果实的呼吸强度、乙烯释放量及多聚半乳糖醛酸酶(PG)活性进行了测定和分析。结果表明:具rin、nor、alc基因的番茄果实无呼吸和乙烯跃变期,其硬度降低慢,果实中PG活性较低,它们的贮藏指数是正常成熟番茄的3倍,一般可贮藏2～3个月,其与正常番茄亲本品种交配的F_1杂种果实(rin/+,nor/+,alc/+)均有一小的呼吸、乙烯高峰出现,但跃变期比正常番茄推迟5～7天,达跃变期时,果实释放的CO_2和乙烯量是正常番茄的40％～6％,PG活性和硬度变化介于双亲之间,果色与正常成熟番茄相近,果实的耐贮藏性较正常番茄有所提高。     

PCR-based differentiation of Fusarium oxysporum ff. sp. lycopersici and radicis-lycopersici and races of F. oxysporum f. sp. lycopersici

The pathogenic type (form and race) of Fusarium oxysporum, which generates wilt symptoms on tomato, was rapidly identified with a polymerase chain reaction (PCR)-based technique. We compared the partial nucleotide sequences of endo polygalacturonase (pg1) and exo polygalacturonase (pgx4) genes from isolates of F. oxysporum ff. sp. lycopersici (FOL) and radicis-lycopersici (FORL) from Japan and designed specific primer sets (uni, sp13, sp23, and sprl) based on the nucleotide differences that appeared among the pathogenic types. PCR with the uni primer set amplified a 670∼672-bp fragment from all isolates of FOL and FORL. With the sp13 primer set, an amplicon of 445 bp was obtained only from isolates of FOL race 1 and 3. With the sp23 primer set, a 518-bp fragment was obtained from isolates of FOL race 2 and 3. The sprl primer set yielded a 947-bp fragment from isolates of FORL, but not from FOL. A combination of amplifications with these primer sets effectively differentiated the pathogenic types of F. oxysporum in tomato.     

3类生境下脐橙果实细胞壁酶、瓤囊壁超微结构和品质变化的研究

为改进脐橙果实质地和科学生态区划与高品质栽培技术提供理论依据,研究了四川3类不同生境下脐橙果实果胶甲酯酶(PE)、多聚半乳糖醛酸酶(PG)、纤维素酶(CX)、瓤囊壁超微结构及主要品质指标的变化。结果表明:果实生长发育期细胞壁酶活性逐渐升高,但不同生境类型的变化趋势不同。超微结构中,南亚热带攀枝花脐橙的线粒体明显,中胶层清晰,细胞壁形态结构较完整;中亚热带江安和中北亚热带雅安脐橙细胞壁的中胶层明显裂解,线粒体膜分解,细胞器空泡化。不同生境SSC和可溶性糖差异显著。PG酶活性与≥10℃积温、1月份平均气温、年平均气温显著负相关,与年降雨量显著正相关。固酸比与≥10℃积温、年平均气温、1月均气温极显著正相关,与日照时数极显著正相关,与降雨量显著负相关。     

大豆疫霉菌多聚半乳糖醛酸酶pspg1基因的克隆及表达分析

大豆疫病严重影响我同及世界各国的农业生产,为探讨多聚半乳糖醛酸酶在大豆疫霉菌致病过程中的作用,采用PCR的方法从大豆疫霉菌中克隆了多聚半乳糖醛酸酶pspg1基因,并利用RT-PCR法对其在大豆中的表达进行了分析.结果表明:大豆疫霉菌pspg1基因开放阅读框长1236 bp,编码一个长412氨基酸的蛋白质.对其进化关系进行分析,发现该基因与其它卵菌的pg基因亲缘关系最近,形成一个独立的分支.RT-PCR分析表明:pspg1基因在接种大豆疫霉菌的大豆下胚轴中大量表达,而在健康大豆下胚轴中未检测到.克隆了大豆疫霉菌pspg1基因,并发现该基因在大豆疫霉菌侵染大豆过程中发挥重要作用.     

Effect of efficient pectinolytic bacterial isolates on retting and fibre quality of jute

Four bacterial strains with high balanced polygalacturonase (PG) and pectin lyase (PNL), and xylanase with minimal cellulase activity were used in jute retting. The 16S rDNA analysis revealed that the organisms were: Bacillus sp. L6 (GQ891097), Bacillus pumilus strain EK-17 (GQ891098), B. pumilus strain Geo-03-422 (GQ891103) and B. pumilus strain IK-MB12-518F (GQ891105). The microorganisms in different combinations of consortia showed synergistic effect resulting in increased PG (35.52-46.61 IU/g cell wet weight), PNL (39.79-72.12 U/ml), xylanase (0.705-0.840 μmol/ml/min) and little cellulase (0-0.153 μmol/ml/min) activities. The organisms in different combinations of consortia reduced the retting period from 11 to 13 days as compared to 19 days in the control. Microbial inoculation produced remarkable improvement in jute fibre strength (26.62-28.91 g/tex) and fineness (2.76-2.92 tex) over control. The pH of the post-retting water samples became acidic, and the electrical conductivity (Ec), chemical oxygen demand (COD) and hardness increased. The organisms could be adopted in industrial application for extraction of jute fibre.     

桃PG基因启动子克隆及序列分析

为获得桃多聚半乳糖醛酸酶( PG)基因启动子并分析调控元件,以桃品种大久保基因组DNA为模板,采用染色体步移技术克隆了PG基因的部分序列及其5′侧翼区,并分析其调控元件组成。结果表明,克隆产物长986 bp,3′侧翼序列与 GenBank中的PG基因序列的5′端部分序列同源性为96%,登录号为FJ940722。在789~839 bp位置存在基础启动子序列,转录起始位点位于828 bp的碱基C,其可能性为0.98。除含有CAAT-Box、TATA-Box等基本启动子元件外,还存在众多参与光响应元件、干旱诱导MYB结合位点、水杨酸响应元件等应答激素和胁迫信号元件,表明PG基因的转录和表达可能受到激素、干旱、光照等外界环境的胁迫以及衰老的调控。     

Biochemical and Nutritional Characterization of Three Prickly Pear Species with Different Ripening Behavior

Biochemical and nutritional changes were studied during the ripening process of three Opuntia morphospecies with different ripening behavior: Naranjona (O. ficus-indica), Blanca Cristalina (Opuntia sp.), and Esmeralda (Opuntia sp.) of early, early-intermediate, and intermediate-late ripening, respectively. In loss of fresh weight, Naranjona showed the highest values, while in Blanca Cristalina and Esmeralda, a discrete weight loss was found. No significant differences were found among morphospecies in soluble solids, total titratable acidity and pH during the postharvest days. Blanca Cristalina and Esmeralda showed an increase in the content of carotenoids, while these diminished in Naranjona. The cell wall enzymes evaluated showed particular behaviors during the ripening of each morphospecies suggesting a fine biochemical control and not a clear relationship between fruit softening and enzyme activity. This study provides basic information on prickly pear ripening, in order to understand this process for its control and for improving shelf life.     

油菜cDNA表达文库构建及核盘菌致病因子互作蛋白的筛选和鉴定

核盘菌 [Sclerotinia sclerotiorum (Lib.) de Bary] 是引起油菜菌核病的病原真菌。核盘菌侵染植物细胞早期,可分泌一些细胞壁降解酶, 如不同类型的多聚半乳糖醛酸酶(polygalacturonase, PG)。这些酶对病菌在植物组织上的定植起关键作用, 同时酶活性的高低也决定病原菌的毒性或致病性。PG可以激活细胞的防卫反应而与其酶活性无关。为了解PG的信号转导途径, 利用RT-PCR方法克隆了核盘菌的PG基因, 将PG的编码区与酵母GAL4的DNA结合功能区融合, 构建到酵母诱饵蛋白表达载体PGBKT7中; 然后在酵母中构建了油菜cDNA表达文库。通过酵母双杂交方法在油菜cDNA表达文库中对与PG互作的蛋白进行了筛选, 分离到一个与PG互作的蛋白, 测序及BLAST分析表明该蛋白含有一个与钙离子结合的结构域, 称为C2结构域(C2-domain), 推测该蛋白是一种钙依赖的膜结合蛋白。该蛋白与拟南芥中一个含C2结构域功能未知蛋白的氨基酸同源性高达80.24%。利用半定量PCR分析了核盘菌诱导后该基因在油菜花、茎、叶中的表达, 结果表明该基因在叶片中表达量最高。克隆到的C2蛋白与其他物种中的C2蛋白比较发现, 与钙离子结合的天冬氨酸残基很保守。     

Multivariate analysis of fresh-cut carambola slices stored under different temperatures

Quality of fresh-cut carambola (Averrhoa carambola L.) is related to many chemical and biochemical variables especially those involved with softening and browning, both influenced by storage temperature. To study these effects, a multivariate analysis was used to evaluate slices packaged in vacuum-sealed polyolefin bags, and stored at 2.5 °C, 5 °C and 10 °C, for up to 16 d. The quality of slices at each temperature was correlated with the duration of storage, O₂ and CO₂ concentration in the package, physical chemical constituents, and activity of enzymes involved in softening (PG) and browning (PPO) metabolism. Three quality groups were identified by hierarchical cluster analysis, and the classification of the components within each of these groups was obtained from a principal component analysis (PCA). The characterization of samples by PCA clearly distinguished acceptable and non-acceptable slices. According to PCA, acceptable slices presented higher ascorbic acid content, greater hue angles (°h) and final lightness (L-5) in the first principal component (PC1). On the other hand, non-acceptable slices presented higher total pectin content, PPO activity in the PC1. Non-acceptable slices also presented higher soluble pectin content, increased pectin solubilisation and higher CO₂ concentration in the second principal component (PC2) whereas acceptable slices showed lower total sugar content. The hierarchical cluster and PCA analyses were useful for discriminating the quality of slices stored at different temperatures.     

多聚半乳糖醛酸酶基因家族的分子进化

多聚半乳糖醛酸酶(polygalacturonase,PG)广泛存在于细菌、真菌和植物中,目前数据库中已积累了大量的序列资料.为了进一步了解PG基因家族的分子进化,以及其作为系统进化研究材料的可能性,本研究选取了88条来自不同物种包括细菌、真菌和植物的PG蛋白序列,用CLUSTALW程序对其氨基酸序列进行了联配,并用邻位相接法构建了系统进化树.结果表明,尽管不同来源的PG序列表现了很大的差异,但仍有4个保守性极强的结构域存在(~(175)NTD、~(197)DD、~(218)GHG和~(250)RIK);同时发现在PG序列中大多数的Cys 位点均具有较好的保守性,而且在细菌、真菌和植物的PG序列中各有自身保守的Cys位点,Cys的保守性体现了物种的分类关系;进一步构建分子进化树显示,PG基因基本上首先根据物种类型而聚类,明显地划分为细菌、真菌和植物三大类;其次,不同作用方式的内切PG(endo-PG)和外切PG(exo-PG)在各类群中也都处于完全独立的分支中.