A single amino acid substitution in the SdhB protein of succinate dehydrogenase determines resistance to amicarthiazol in Xanthomonas oryzae pv. oryzae

BACKGROUND: Xanthomonas oryzae pv. Oryzae Ishiyama, a causal agent of rice bacterial leaf blight, was found to be sensitive in vitro to the systemic fungicide amicarthiazol (2‐amino‐4‐methylthiazole ‐5‐carboxanilide), which is a potent inhibitor of succinate dehydrogenase (SDH, EC 1.3.99.1). This paper aimed to determine the molecular resistance mechanism of X. oryzae pv. oryzae to amicarthiazol. RESULTS: UV‐induced resistant mutants of X. oryzae pv. oryzae to amicarthiazol were isolated. The activity of SDH in wild‐type X. oryzae pv. oryzae was strongly inhibited by amicarthiazol, while that in resistant mutants was insensitive, although their SDH activity was decreased compared with the wild‐type sensitive strain without amicarthiazol. A mutation of Histidine₂₂₉ (CAC) to Tyrosine₂₂₉ (TAC) was identified in sdhB, which encoded the iron–sulfur protein subunit of SDH. The sdhB from the mutant was ligated into a cosmid, pUFR034, to generate pUFR034RAX, which conferred resistance to amicarthiazol when transformed into the wild‐type sensitive strain. CONCLUSION: A mutation of His₂₂₉ (CAC) to Tyr₂₂₉ (TAC) in SdhB was responsible for determining amicarthiazol resistance. Copyright © 2010 Society of Chemical Industry     

酚类化合物对发光细菌的急性毒性和对ETS的抑制研究

研究了5种酚类化合物对发光细菌毒性和活性污泥脱氢酶活性的抑制作用。结果表明,酚类化合物对502海水发光细菌的毒性大小顺序为2,4-二氯苯酚>对硝基苯酚>邻甲基苯酚>苯酚>对氨基苯酚,对应的EC50值分别为4.95、23.90、42.43、83.65和178.75 mg/L。而对活性污泥脱氢酶活性抑制的大小顺序同发光细菌毒性测定结果不太一致,毒性大小顺序为2,4-二氯苯酚>邻甲基苯酚>对氨基苯酚>苯酚>对硝基苯酚,相应的EC50值分别为37、398、575、794和1 333 mg/L。两种毒性测定方法的EC50结果差异比较大,可能和测定方法的特点有关。但仅仅根据单一的、高灵敏度的发光细菌毒性测定结果来调整废水处理工艺中的各项参数而实现对水质的调控,可能会造成处理成本的大幅度地增加,因此根据两种或多种毒性测定方法(其中包括以活性污泥为对象)的测定结果判定有机污染物的毒性更科学。     

昆明裂腹鱼不同组织乳酸脱氢酶同工酶研究

采用不连续聚丙烯酰胺凝胶电泳法,对昆明裂腹鱼的眼球、心脏、肾脏、性腺、脾脏、肝胰脏、脑、鳃和肌肉等9种组织的乳酸脱氢酶同工酶(LDH)进行了测定。结果表明:昆明裂腹鱼LDH共有15条酶带表现出特异性,在不同组织中检出的LDH酶带数量不同,以肝胰脏中最多,共检出9条;在酶活性方面,LDH在脑、鳃、肌肉和肾脏等组织中活性最强,而在眼球组织中最弱。     

擎天凤梨GAPDH基因的克隆及序列分析

管家基因GAPDH(glyceraldehyde-3-phosphate dehydrogenase)常被用作定量、半定量PCR试验的内参,以确定目标基因的相对表达量。基于前期从全长cDNA文库中获得GAPDH基因的EST单克隆,进行Primer Walking测序,获得一个GAPDH的全长cDNA序列,序列长1 790 bp,编码421个蛋白氨基酸,命名为GoGAPDH1。采用Blast,ExPASy,ProtParam,SPOMA,Find Conserved Domains(NCBI),ClustalX, MEGA等生物信息学软件对cDNA 序列、氨基酸序列、保守区、亲缘关系等特征进行分析。结果表明,GoGAPDH1与玉米的细胞质GAPDH基因源性最高,为82%。初步推断GoGAPDH1可能是一个胞质型GAPDH。     

吲熟酯和钼酸铵处理对青种枇杷果实糖积累及相关酶活性的影响

以青种枇杷为试材,在果实成熟前50d左右分别喷施100、200、300mg·L-1吲熟酯及0.1%钼酸铵和200mg·L-1吲熟酯+0.1%钼酸铵,研究吲熟酯和钼酸铵对果实生长发育过程中糖积累以及蔗糖与山梨醇代谢相关酶活性变化的影响。结果表明:200mg·L-1吲熟酯处理对增加果实的果糖和葡萄糖积累最为明显。所有处理的果实山梨醇含量均随果实发育呈先下降后上升的趋势。100mg·L-1吲熟酯处理的蔗糖合成酶(SS)和蔗糖磷酸合成酶(SPS)活性在5月21日后呈明显的下降趋势,其他处理的蔗糖代谢相关酶活性均呈上升趋势;山梨醇代谢相关酶活性总体上呈下降趋势。综上所述,200mg·L-1吲熟酯处理对青种枇杷果实糖积累有较显著的促进作用。     

复合型丸粒剂对防风种子萌发的影响

研究了复合型丸粒剂对防风种子发芽和发芽生理的影响。结果表明,防风种子经复合型丸粒剂处理后,种子活力、过氧化物酶和脱氢酶活性显著提高。丸粒化防风种子发芽集中度较裸种子显著提高,为裸种子的1.39倍。这说明复合型丸粒剂可有效改善防风种子的萌发。     

乙醛脱氢酶基因（ALDH）转化番茄的研究

乙醛脱氢酶基因（ALDH）为目的基因,构建了pBI-ALDH 植物表达载体,番茄品系03HN31子叶为外植体,经发根农杆菌（Agrobacterium rhizogens）介导采用共培养法转化番茄。通过分子生物学技术（PCR、Southern杂交、RT-PCR）和抗逆相关生理指标测定（相对电导率及丙二醛含量）相结合的方法对转化番茄植株进行检测,研究结果表明：乙醛脱氢酶基因（ ALDH ）导入并整合到番茄的基因组中,在转录水平上可以稳定表达;在干旱、高盐和低温胁迫条件下,转基因番茄植株和对照株的相对电导率和丙二醛含量有差异,证明转化番茄植株的质膜受损程度有所降低,抗氧化胁迫性能有所提高,转化率为10.78%。     

Modulation of manganese toxicity in Pisum sativum L. seedlings by kinetin

In the present study, the effects of kinetin (KN; 10 and 100 μM) application under manganese toxicity (Mn; 50, 100 and 250 μM) were investigated, on growth, photosynthetic pigments, total protein, total nitrogen, ammonium (NH₄⁺) content, NH₄⁺ assimilating enzymes and antioxidant system in pea seedlings. The exposure of pea seedlings to Mn and 100 μM of KN alone and in combination, caused decrease in growth, photosynthetic pigments, total protein and total nitrogen contents, and an increase in NH₄⁺ content. However, application of 10 μM of KN together with Mn reduced the Mn toxicity symptoms, promoted the growth of seedlings and led to the decrease in NH₄⁺ content compared to Mn treatments alone. The root and shoot activities of glutamine synthetase (GS), glutamate oxoglutarate aminotransferase (GOGAT) and catalase (CAT) were decreased while glutathione reductase (GR) and dehydroascorbate reductase (DHAR) activities exhibited differential responses when pea seedlings were exposed to Mn and 100 μM of KN. However, under similar treatments, activities of glutamate dehydrogenase (GDH), superoxide dismutase (SOD) and ascorbate peroxidase (APX) in root and shoot were increased. It was noticed that addition of 10 μM of KN together with Mn, caused significant stimulation in activities of enzymes of NH₄⁺ assimilation and antioxidant defense system even over their respective control values. Non-enzymatic antioxidants (ascorbate and glutathione) in root and shoot of pea seedlings exposed to Mn stress were significantly increased by the addition of 10 μM of KN. Therefore, ameliorative effect of 10 μM of KN against Mn toxicity was observed. This study thus suggests that 10 μM of KN appreciably improves Mn tolerance of pea seedlings under Mn toxicity while reverse effects were exhibited by 100 μM of KN.     

不同施氮处理对水稻颖果灌浆和呼吸活性的影响

为了研究施氮量和施氮时期对水稻颖果生理活性的影响,以一籼一粳两个水稻品种为材料,施用分蘖肥和孕穗肥,并在施氮总量相同的条件下设置了前高后低和前低后高两个处理。研究结果表明：增施氮肥维持了水稻颖果灌浆后期较高的含水率,延长了颖果灌浆充实的时间;维持了水稻颖果灌浆充实期尤其是灌浆后期较高的呼吸速率,并且使颖果维持较高呼吸活性的时间延长;延长了颖果各组织维持脱氢酶活性的时间。其中孕穗肥较高的施氮处理比分蘖肥较高的施氮处理效果更为显著。