用于筛选植物PGIP的酵母双杂交PGase诱饵载体

对真菌抗性较强的植物能利用自身的多聚半乳糖醛酸酶抑制蛋白(po lyga lacturonase-inh ib iting prote in,PG IP)来抑制真菌分泌的、降解植物细胞壁的主要物质——多聚半乳糖醛酸酶(po lyga lacturonase,PG ase)的活性.目前,快速、有效的获取植物PG IP的方法是通过酵母双杂交技术来进行的,而这首先要获得真菌PG ase的酵母诱饵载体.以真菌——互格链格孢PG ase的cDNA的为基础,将其克隆到M ATCHM AKER L exA Tw o-Hybrid System的诱饵载体中,并将诱饵载体(pL exA-PG ase)成功转化酵母菌株EGY 478[p8op-lacZ],经检测无自激活作用,可直接用于快速、大量地筛选植物PG IP.     

Differential response of zucchini varieties to low storage temperature

Several parameters related to cold damage in zucchini have been measured in fruits from different varieties cultivated in the province of Almería (south-eastern Spain). The purpose of this work is to correlate physiological parameters with commercial quality and chilling damage, in order to establish reliable criteria for selecting varieties having better genetic adaptation to low-temperature storage. The results show that, for varieties harvested during February, the variety Natura was the best adapted to storage at 4 °C, as its fruits suffered less from chilling injuries, had a lower weight loss, and the levels of metabolites malonyldialdehyde and H₂O₂ were not as high as in the other varieties. Genetic variation was detected for some of the parameters analysed. The positive correlation among the levels of MDA, H₂O_2, and chilling injury, as well as the negative correlation among catalase and chilling injury, makes these parameters good indicators of chilling damage in our system.     

授粉对度尾文旦柚果皮相关酶活性的影响

研究了利用度柚6号、琯溪蜜柚授粉处理对度尾文旦柚果实发育后期果皮多聚半乳糖醛酸酶(PG)、纤维素酶(CX)及苯丙氨酸裂解酶(PAL)活性的影响。结果表明:授粉处理后,度尾文旦柚果实发育后期PG活性呈上升趋势;CX活性呈下降趋势;PAL活性呈先上升后下降的趋势。     

Maceration of plant tissue by fungi is inhibited by recombinant antipectinase antibodies

Polyclonal antiserum from mice immunized with extracellular proteins from Rhizoctonia solani inhibited pectinase and cellulase activities in cell free culture supernatants of Rhizoctonia solani. Spleen mRNA from these mice was used to construct a cDNA library from which antipectinase ScFv antibodies were isolated using phage display techniques. Soluble ScFv antibodies produced by individual clones in Escherichia coli inhibited polygalacturonase in the culture supernatants of a range of fungal pathogens, including ascomycetes, basidiomycetes, and oomycetes. The soluble antibodies also inhibited maceration of potato tissue by these pathogens.     

Enzymatic Maceration of Witloof Chicory by the Soft Rot Bacteria Erwinia carotovora Subsp. carotovora: The Effect of Nitrogen and Calcium Treatments of the Plant on Pectic Enzyme Production and Disease Development

Disease incidence of bacterial soft rot caused by Erwinia carotovora subsp. carotovora and activities of bacterial pectolytic enzymes were studied in witloof chicory. Disease incidence after forcing of the chicory heads was enhanced by the nitrogen treatment and reduced by the calcium treatment of the chicory plants prior to forcing. Significant differences in susceptibility to bacterial soft rot were found for the tested chicory cultivars. Disease severity after 96h ranged from 6% in Clause R2 to 100% in Bea and Tabor. Chicory cultivars Rumba and Salsa showed a final average severity of 65–70%. Activity of the pectolytic enzymes, polygalacturonase and pectate lyase increased in artificially inoculated chicory heads of cultivar Rumba and was not affected by calcium and nitrogen treatments of the plants. Polygalacturonase showed highest activity 48h after inoculation while pectate lyase activity increased throughout 72h of incubation. Maceration of the chicory tissue and bacterial growth increased continuously until 96h after inoculation, when more than 60% of the chicory heads was macerated by pectolytic enzymes of the bacteria. Enzyme activity of Erwinia carotovora subsp. carotovora grown on chicory cell wall extracts was influenced by nitrogen and calcium treatments of the chicory plants. The activity of polygalacturonase reached its highest level at 48h after incubation and pectate lyase activity peaked at 24h followed by a continuous decrease until 72h after inoculation.     

TDZ处理对棉花叶片脱落率及酶活性的影响

【目的】研究棉花在 TDZ (Thidiazuron, TDZ)处理后叶片脱落的机制。【方法】以中棉所35号、新陆早19号、农大5号、XND1378和新陆早50号5个棉花品种为材料,于2017和2018年在塔城沙湾新疆农业大学棉花基地和塔城奎屯农七师农科所实验地,吐絮期分别设置两个处理 T1(大田喷施 TDZ 时间前4 d)和 T2(与大田处理时间相同), TDZ 处理后,研究脱叶率,根据脱叶率给5个棉花品种排序,于0、6、18、42、66和96 h测定叶托、叶片、叶柄的多聚半乳糖醛酸酶(PG)和纤维素酶活性,统计叶片脱落率、吐絮率和棉花产量。【结果】喷施 TDZ 后,T1的棉花产量显著低于 CK 和 T2 (P＜0.05);两个处理下棉花各材料的脱落率和吐絮率均极显著高于 CK (P＜0.01);叶托的 PG 活性在6、18 、42、66和96 h五个时期显著高于 CK (P＜0.05);纤维素酶活性在6、66 和96 h三个时期显著高于 CK (P＜0.05),在18、42 h两个时期极显著高于 CK (P＜0.01)。【结论】中棉所35号和新陆早50号对TDZ最敏感,TDZ处理可调节脱落叶片的 PG 活性和纤维素酶活性,提高叶片脱落率,有利于棉花机械采收。     

Activity of polygalacturonases from Moniliophthora perniciosa depends on fungus culture conditions and is enhanced by Theobroma cacao extracts

We report the first analysis of polygalacturonase regulation in the basidiomycete Moniliophthora perniciosa. Non-secreted and secreted polygalacturonase activity was obtained from M. perniciosa cultivated on bran-based solid medium or liquid media containing additional carbon sources or cacao extracts (infected or not by the fungus). Polygalacturonase activity assays were carried out under different temperatures and incubation periods. The best secreted polygalacturonase activity was obtained when the enzymatic assay was made at 50 °C for 10 min. Moreover, the polygalacturonase activity was enhanced when the fungus was cultivated on potato dextrose medium, in the presence of additional fermentable carbon sources, in the presence of cacao pulp or non infected cacao extracts.     

冷激对贮藏白果多聚半乳糖醛酸酶的影响

以白果 (Ginkgo bilba L.)为材料 ,以 0℃冰水混合物为冷却介质 ,研究了不同冷激处理对贮藏期间多聚半乳糖醛酸酶 (PG)活性的影响 .结果表明 ,冷激能显著抑制 PG活性 ,抑制程度随冷激处理时间的延长而加强 .     

大豆疫霉根腐病菌多聚半乳糖醛酸酶活性对其致病力的影响

就大豆疫霉根腐病菌（Phytophthora sojae）多聚半乳糖醛酸酶（Polygalacturonase,PG）活性对其致病力的影响进行了研究。结果表明,受试致病性菌株在接种复壮后PG活性明显高于复壮前的PG活性;致病性菌株的PG活性显著高于非致病性菌株PG活性;受试菌株的PG活性均在培养后第7天达到峰值。综上所述,PG活性的高低与大豆疫霉根腐病菌的致病力强弱有一定的关系,是该病菌的一种起致病作用的细胞壁降解酶。     

果实软化相关PG基因的进化分析和基因组定位

 利用NCBI数据库和蔷薇科基因组数据库中的数据对48个果实PG基因进行系统进化分析,结果表明3个进化支中都包含与果实软化相关的PG基因,属于A支和属于B支的PG基因有明显的功能差异。氨基酸序列分析显示,3个进化支中的PG基因存在多个差异位点,A支和C支PG基因的N端比B支短60 ~ 70个氨基酸。PG基因结构（包括启动子、内含子和编码区）的变异对其功能差异有重要影响。对苹果和桃的PG基因进行了基因组精确定位,其附近存在果实软化相关性状标记。