浅议等电聚焦电泳技术在种子纯度检测中的应用

结合多年试验从制胶方法、凝胶浓度、样品提取、染色液的储备及染色方法等方面总结了利用酯酶、过氧化物酶、磷酸葡萄糖变位酶、苹果酸脱氢酶等同工酶谱带鉴别大白菜、黄瓜、番茄、甘蓝等多种蔬菜几十个品种的种子纯度,综述了等电聚焦电泳技术在种子纯度检测中的应用。     

外源水杨酸对冷藏鸭梨呼吸途径及关键酶的影响

以鸭梨为试材,研究不同浓度的外源水杨酸对梨果实采后冷藏的呼吸途径及呼吸关键酶的影响。研究结果表明：水杨酸降低了糖酵解途径（EMP）、三羧酸循环途径（TCA）、磷酸戊糖途径（PPP）和细胞色素途径、交替途径的表达,降低了磷酸己糖异构酶、琥珀酸脱氢酶、细胞色素氧化酶、葡萄糖．6磷酸脱氢酶和6．磷酸葡萄糖酸脱氢酶联合活性及总...     

墨兰、春兰变种和品种间同工酶分析

 通过酯酶( EST) 、苹果酸酶(ME) 、磷酸葡萄糖异构酶;PGI ( NADP) 、磷酸葡萄糖变位酶PGM( NAD) 、超氧化物歧化酶( SOD) 5 种酶系统研究墨兰和春兰17 个变种及品种的同工酶多样性, 并探讨这些材料间的亲缘关系。5 种酶系统所得的11 个位点中7 个是多态性位点。EST、PGM( NAD) 和SOD具有多态性。上述5 种酶系统能够把供试品种和变种区分开, 并划分为墨兰和春兰类。     

介绍一种快速测定辣椒异交率的方法

Stevern报道了一种同工酶进行辣椒自然异交率(NCP)的测定方法,简单易行。现介绍如下: 辣椒的种子中含有一种2一磷酸葡萄糖酸变位酶(Pgm-2),它在一些辣椒系统如PI164502, PI 174810,PI 174112, PI 138567的纯合个体中表现出慢的同工酶谱带,而在另外一些系统如美国广泛栽培的品种     

粤北野生灵芝与栽培灵芝同工酶及可溶性蛋白的研究

采用垂直聚丙烯酰胺凝胶电泳技术,分别对粤北5株野生灵芝与9株栽培灵芝进行酯酶和过氧化物酶同工酶及可溶性蛋白进行比较与分析,并通过聚类分析对比各菌株间的遗传差异性.结果表明:酯酶和过氧化物酶同工酶酶谱在酶带数量和酶活性方面都表现出一定的差异;供试各菌株间可溶性蛋白差别不大.进一步采用两种同工酶综合聚类分析发现,14株菌株分为13类,野生灵芝菌株与栽培菌株之间差异较大;这也表明同工酶技术在灵芝分类、鉴定、和遗传标记等方面都具有参考作用,且粤北地区野生灵芝资源潜在较大的开发利用价值.     

从生物化学的角度看担子菌子实体形成

总结了从生物化学的角度看待担子菌子实体形成的研究进展。通过以金针菇为模型的RI示踪实验,解析了菌丝体中积蓄的糖原作为营养成长过程中的主要贮藏碳水化合物、随着子实体形成的开始而被分解、被转换为海藻糖后输送到子实体、并作为主要发育基质被利用。笔者率先发现了存在于海藻糖的合成与分解代谢中的新酶—海藻糖磷酸化酶(TP),接着,在几种食用菌的生化反应中证明了此酶存在并通过基因克隆方法发现此酶在子实体形成中的作用—在营养菌丝体中以糖原为基质催化加磷酸反应,形成α-G1P海藻糖,当海藻糖输送到子实体,借助该酶的加磷酸分解功能生成α-G1P葡萄糖,而葡萄糖作为子实体发育中的主要糖代谢基质而被加以利用。了解了营养菌丝从培养基中吸收的葡萄糖基本上不能从菌丝体输送到子实体。通过基因实验,检证了TP在营养菌丝体中对催化海藻糖的合成作用。另一方面,该酶还具有加磷酸分解从菌丝体输送到子实体的海藻糖的生理功能,伴随子实体发育的细胞内磷酸糖类分析证明了α-G1P(α-1磷酸葡萄糖)的含量比α-G6P(α-6磷酸葡萄糖)更多。另外,从子实体形成过程中氮化合物代谢分析表明,营养菌丝的菌体蛋白为形成子实体的主要氮素来源,当生殖生长启动时被菌丝体内的蛋白酶分解,形成的氨基酸输送到子实体并被用于子实体发育。通过这些研究的累积,已大致了解了碳素和氮素在子实体形成过程中的动态,但是,从营养生长转换到子实体形成的机制却至今仍不明了。     

^60Coγ射线辐照对东方百合不定芽POD同工酶的影响

运用60Coγ射线辐照东方百合离体不定芽,对不定芽不同生长时期组织过氧化物(POD)同工酶酶谱进行分析,结果表明:辐照后POD同工酶酶带数和酶带颜色与对照比较都存在一定差异,两个时期6.0、8.0Gy处理的POD同工酶均出现了一条低分子量酶带,对同工酶进行聚类分析发现高剂量处理(≥4.0 Gy)与对照的相似性较小,差异较大.     

水菜二种同工酶指纹图谱及分类研究

同工酶酶谱分析和聚类分析表明,水菜属于芸薹种,可将水菜作为亚种,京水菜和壬生菜作为水菜亚种下的变种处理.PPO同工酶用于芸薹属种间的分类鉴定优于POD同工酶.叶PPO同工酶E'4为水菜的特异酶带.     

茶薪菇不同生长期同工酶的研究

本文报道了茶薪菇Ａgrocybe Chaxingu不同生长期的氧化物酶（ＰＯＤ）、酯酶、（ＥＳＴ）超氧物歧化酶（ＳＯＤ）的同工酶谱分析及三种同工酶谱聚类分析树状图。结果表明，茶薪菇菌丝体与子实体同工酶存在着明显差异，二者的相似水平只有４５％。     

生化和ISSR分子标记在黑木耳品种鉴定中的应用

以生产收集的24个黑木耳栽培菌株为试材,采用拮抗、酯酶同工酶及ISSR分子标记技术对其进行分类鉴定和遗传多样性分析。结果表明:酯酶同工酶酶谱显示24个黑木耳菌株共扩增出11条迁移率不同的酶带,聚类分析表明当遗传系数为0.74时,将24个黑木耳菌株分为两大类;通过ISSR分子标记共扩增出67条清晰的DNA多态片段,大小介于0.2～2.0 kb,聚类分析表明当遗传系数为0.85时,将24个菌株分为两大类,聚类分析结果与拮抗试验结果基本一致。该研究为黑木耳的菌株选择及遗传育种提供了参考依据。     

两个柑桔杂交组合F_1代的几种同工酶的遗传分析

采用聚丙烯酰胺凝胶电泳对柑桔两个杂交组合克里迈丁×三湖化红、克里迈丁×宜昌橙F_1代杂种各30侏的GOT、SOD、POX、ME四种酶的同工酶进行了检测分析,发现在GOT—2、SOD—2、POX—2、ME—2四个酶位点上F_1代有分离,表明亲本在这些酶基因位点上处于杂合状态。GOT—2、SOD—2、POX—2的分离经x~2测验,符合孟德尔单基因遗传,与其相应的Got—2、Sod—2、Pox—2三个酶基因可作为遗传标记应用。ME—2的分离复杂,说明柑桔在遗传基础上十分复杂。     

Identification and investigation of relationships of mandarin types using isozyme analysis

Sixteen enzyme systems with 19 loci were used to discriminate 19 mandarin cultivars and hybrids. Variability was observed at 12 loci, and all but 3 mandarin types could be discriminated, 2 of which were probably identical. Relatedness within the tangelo and tangor groups was high, probably reflecting their recent origin. Relatedness within the common mandarin group was low, reflecting their multiple origins and long cultivation. Exceptions were cultivars ‘Algerian’ and ‘Beauty of Glen Retreat’, which differed at only 1 locus. Relatedness between the groups was generally low, with the least relatedness between the tangelos and the other groups, probably owing to their grapefruit parent. The ‘Ellendale’ cultivars formed a particularly cohesive group, but with 2 isozyme genotypes, differing at 2 loci, both marketed as ‘Ellendale’. Three of the ‘Ellendale’-type cultivars probably arose by self-pollination of ‘Ellendale’, the rest either by self-pollination or by mutation.     

不同比例平菇废料栽培鸡腿蘑子实体的同工酶研究

不同比例平菇废料栽培鸡腿蘑子实体的同工酶分析结果表明,６个参试的鸡腿蘑样品过氧化物酶（ＰＯＤ）同工酶谱差异明显,而其酯酶（ＥＳＴ）和超氧化物歧化酶（ＳＯＤ）同工酶谱基本相同。     

Plant growth,yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on four rootstocks

Vegetative growth, yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on ‘Rangpur’ lime, ‘Swingle’ citrumelo, ‘Orlando’ tangelo, and ‘Cleopatra’ mandarin were evaluated under subtropical climate of Northern São Paulo State, Brazil, from 2000 through 2006 harvest seasons. ‘Fallglo’ mandarin trees had the highest cumulative yield on ‘Rangpur’ lime, and the smallest on ‘Swingle’ citrumelo and ‘Orlando’ tangelo. Plants of this cultivar had the highest yield efficiency on ‘Rangpur’ lime, and the lowest on ‘Orlando’ tangelo. ‘Sunburst’ mandarin trees began to bear fruits later than ‘Fallglo’ mandarin trees, with no differences in yield induced by the rootstocks. ‘Cleopatra’ mandarin induced the most vigorous growth in ‘Fallglo’ mandarin as compared to plants on ‘Swingle’ citrumelo. On the other hand, the largest trees of ‘Sunburst’ mandarin were registered on ‘Orlando’ tangelo, and the smallest on ‘Rangpur’ lime. ‘Sunburst’ mandarin had higher alternate bearing than ‘Fallglo’ regardless the rootstock. Fruit weight and juice content were not affected by the rootstock. These two mandarin scion cultivars may be considered adequate alternatives to produce good fruit quality for the fresh fruit market. ‘Cleopatra’ mandarin and ‘Rangpur lime are suitable rootstocks for ‘Fallglo’ mandarin, whereas all rootstocks evaluated are adequate for ‘Sunburst’ mandarin.     

孔雀草POD同工酶及主要观赏性状研究

采用垂直平板聚丙烯酰胺凝胶电泳(PAGE)方法对6个孔雀草品种的过氧化物酶(POD)同工酶进行了研究,并观测了其主要观赏性状,初步探讨了两者间的关系.结果表明:6个孔雀草品种间的POD同工酶酶谱存在不同程度的差异,同一品种不同器官的POD同工酶间存在明显的差异;分析了6个品种的主要观赏性状,雪域的综合表现最好;主要观赏性状与POD同工酶酶谱间未表现出明显的规律性的关系.     

Evaluation of ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid for tolerance to Xanthomonas axonopodis pv. citri and Xylella fastidiosa

Asiatic citrus canker (ACC), caused by Xanthomonas axonopodis Starr & Garces pv. citri (Hasse) Vauterin et al., and citrus variegated chlorosis (CVC), caused by Xylella fastidiosa Wells et al., are considered the main diseases affecting sweet orange scion varieties in Brazil. Among commercial varieties, mandarins and tangerines are recognized as tolerant to these pathogens. We report herein the production of ‘Hamlin’ sweet orange (Citrus sinensis L. Osbeck) + ‘Montenegrina’ mandarin (Citrus deliciosa Ten.) allotetraploid somatic hybrid plants by protoplast fusion with improved disease tolerance that could be used as a donor of resistance genes in interploid hybridisation. Somatic hybridisation was confirmed by leaf morphology, flow cytometry and RAPD analyses. The somatic hybrid was propagated by grafting and cultivated in a screenhouse for tolerance assays. For X. axonopodis pv. citri assays, buds were collected from both ‘Hamlin’ sweet orange and the somatic hybrid and grafted onto ‘Cleopatra’ mandarin (Citrus reshni hort. ex Tanaka). As a negative control, buds from ‘Mexerica Tardia’ mandarin (C. deliciosa) were collected and grafted onto ‘Cleopatra’ mandarin. Two-month old plants with at least one young vegetative flush were individually spray-inoculated with a 10⁶ CFU mL⁻¹X. axonopodis pv. citri suspension and incubated in a growth chamber, at 27 °C, under 16-h photoperiod. The somatic hybrid showed a statistically significant reduction in susceptibility to ACC 30 days after inoculation. Compared to ‘Hamlin’ sweet orange, disease severity was reduced by 70%, with similar tolerance to that of the mandarin negative control. For X. fastidiosa assays, buds were collected from the somatic hybrid and its parental plants and grafted onto ‘Rangpur’ lime (Citrus limonia Osbeck). The developed plants were needle-inoculated with a X. fastidiosa suspension (8.7 × 10¹⁰ CFU mL⁻¹) into the new growth flush stem. Bacterial population was quantified both at 4 (at the inoculation point) and 8 months (50 cm above the inoculation point) after inoculation. The first evaluation detected X. fastidiosa in 63% of ‘Hamlin’ sweet orange and ‘Hamlin’ + ‘Montenegrina’ mandarin samples. In the second evaluation, X. fastidiosa was detected in 47.4% of ‘Hamlin’ sweet orange and 10.5% of ‘Hamlin’ + ‘Montenegrina’ somatic hybrid samples, suggesting that bacterial movement was restricted in the somatic hybrid. X. fastidiosa was not detected in both evaluations in samples collected from leaves of ‘Montenegrina’ mandarin. These results indicate that the ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid has potential for improved disease tolerance that should enhance its value regarding future use in citrus breeding programs.     

软枣猕猴桃植株的过氧化物酶同工酶酶谱差异研究

对软枣猕猴桃7个样品的过氧化物酶同工酶酶谱的差异进行了分析。结果表明:同工酶标记对软枣猕猴桃幼苗性别鉴定是有帮助的,但还需要继续加深此类研究才能更精确地进行幼苗性别鉴定,并对今后的研究重点和解决办法提出了建议。     

宽皮柑橘单核苷酸多态性的高分辨率熔解曲线分型

 高分辨率熔解曲线分析（High resolution melting analysis,HRM）可以检测单碱基改变引起的DNA双链熔解温度（T_m）值变化,从而可以对样本在单核苷酸多态性分子标记（Single nucleotide polymor- phism,SNP）上进行基因分型。通过分析NCBI数据库中宽皮柑橘的表达序列标签（Expressed sequence tag,EST）数据鉴别SNP位点,并用小片段扩增法高分辨率熔解曲线分型技术（High resolution melting analysis of small amplicons）分析11个宽皮柑橘（Citrus reticulata）品种以及柳橙（Citrus sinensis Osbeck var.‘Liucheng’）的5个SNP位点的基因型。结果显示,小片段扩增法高分辨率熔解曲线分型可以快速、清楚地分辨纯合与杂合基因型,在校正温度差异后也可以很好地分辨同一个SNP位点不同的纯合型。统计分析表明样本在所有SNP位点上均存在多态性,5个SNP位点的平均多态性信息含量（PIC）为0.3190,显示样本在这组SNP位点上具有较高的杂合率。     

两种砧木对砂糖橘磷素累积消耗量的影响

采用以酸橘、枳为砧木的成年砂糖橘树作为供试材料,通过对各器官各部位磷素营养水平的分析,研究磷素累积消耗的特点与规律。结果表明：1、砂糖橘不同器官的磷素累积消耗量存在差异,以枝条的累积消耗量最大;除枝条外,不同砧木砂糖橘的相同器官中磷素累积消耗量差异不显著。2、两种砧木砂糖橘地上部的磷素累积消耗量均显著地高于地下部。3、砂糖橘地上部和地下部各器官不同部位的磷素累积消耗量存在一定差异,但不同砧木对地上部和地下部器官各部位的磷素累积消耗量影响不大。4、砂糖橘各级侧枝木质部与韧皮部的磷素累积消耗量存在一定差异,不同砧木砂糖橘之间也不相同。