根癌农杆菌介导ACS反义基因转化香蕉的研究

以香蕉(Musa spp.)品种‘天宝蕉’(Musa spp.,AAA类群)为试材,对以根癌农杆菌介导法的香蕉遗传转化体系进行较全面的研究,并以该系统进行了ACS反义基因转化香蕉的研究。结果表明:不经预培养的香蕉茎尖横切薄片,侵染前用附加0.1 mg/L甘露醇的高渗固体培养基前处理4 h,农杆菌重悬液浓度为OD600在1.0左右,重悬液中含100 g/L蔗糖,接菌时间为10~15 min,于26℃黑暗条件下共培养4 d,共培养培养基pH值为5.8是较为适合的转化条件;采用附加100 mg/L卡那霉素、2 mg/L AgNO3筛选培养基对共培养后的香蕉横切薄片进行筛选,共获得5个转ACS反义基因的抗性芽系;经GUS组织化学法及PCR检测,gus基因已整合进香蕉基因组。     

甜瓜果实中芳香物质的研究现状

芳香物质是衡量甜瓜果实品质高低的重要指标之一，它是由不同挥发性物质组成的混合物，主要包括醇类、醛类、酮类、萜类、酯类以及含硫化合物等。主要综述不同品种甜瓜芳香物种类及其芳香物相关基因的研究现状。     

乙烯生物合成关键酶基因研究进展

ACC合成酶(ACS)和ACC氧化酶(ACO)是催化乙烯生物合成的两个关键酶，这两个酶的基因在调控果品和花卉完熟及衰老方面具有重要意义，本文综述了ACS和ACO基因最新研究进展。     

5种植物花瓣ACS基因的克隆与分析

分析已发表的植物1-氨基环丙烷-1-羧酸（ACC）合成酶（ACS）基因序列,设计一对简并引物,在随机选择的非洲菊、月季、桂花、茶梅和山茶等5种植物花瓣cDNA中进行PCR扩增．结果表明,在5种植物花瓣中都能扩增出约800bp的特异片段,登录GenBank发现均未被注册,因此将序列提交到GenBank并获得了序列号．在NCBI上进行Blast比对,发现所克隆的5个Acs基因均与其他植物花瓣的ACS基因有一定的序列相似性,最高达94％,最低为80％．将5个ACS基因的片段进行多序列比较分析,发现茶梅ACS基因与山茶ACS基因的序列差异最小,序列相似性达99％．本试验设计的ACS基因简并引物在植物中有一定的通用性,后续可以适当调整引物的简并度,以得到高通用性的简并引物．     

Knockdown of ACS9 expression in Arabidopsis decreases the tolerance to salt and osmotic stress

Based on the DNA sequence of ACS9, two produced fragments were subcloned into binary vector pCAMBIA1300 in antisense and sense orientations, and the generated RNA interference (RNAi) vector was then transformed into Arabidopsis thaliana. The stress resistance function of ACS9 gene in Arabidopsis thaliana was researched by determination of stress resistance physiologic indexes, NaCl and PEG6000 resistance. The results showed that the inhibition of ACS9 expression enhanced the sensitivity to high concentration NaCl (150 mmol/L) and PEG6000 (7%) in Arabidopsis thaliana seeding stage. The proline contents and water loss rates in transgenic plants were 0.68 and 1.4 times higher than those in the wild-type leaves, respectively, indicating that the inhibition of ACS9 expression due to salt and drought resistant was reduced and suggested that ACS9 gene played important roles in plant salt and drought tolerance.     

Evaluation of Associative Effects of Alfalfa Meal and Ammoniated Corn Straw by Gas Production Technique in vitro

This experiment was conducted to determine the best roughage combination of alfalfa meal (AM) and ammoniated corn straw (ACS),so as to improve the efficiency of roughage utilization and reduce the cost of feeding. The AM was mixed with ACS in the proportion of 100:0,80:20,60:40,50:50,40:60,20:80 and 0:100 with 3 replicates per group. The accumulated gas production (GP) at fermentation for 3,6,12,24,48 h,and the changes of pH,dry matter disappearance rate (DMD),ammonia nitrogen (NH₃-N) concentration, microbial protein (MCP) and volatile fat acids (VFA) concentration,and their single factor associative effects index (SFAEI) and multiply factors associative effects index (MFAEI) were determined by gas production technique in vitro after fermentation for 48 h. The results showed as follows:The accumulated gas production of AM20:ACS80 group was higher than that of the other groups,and the DMD of AM0:ACS100 group was significantly lower than other groups (P<0.05).The NH₃-N concentration of AM20:ACS80 and AM0:ACS100 groups was significantly higher than the other groups (P<0.05).The MCP concentration was highest in AM20:ACS80 group,and the concentration of acetic acid and TVFA in AM20:ACS80 group was the highest (P<0.05),and the acetic acid/propionic acid ratio in all groups was more than 3,which indicating that it belonged to the acetic acid fermentation type.The pH during the fermentation ranged from 6.69 to 6.85.The results of MFAEI and SFAEI of each combination indicated that only the AM20:ACS80 group showed the positive associative effects. In conclusion,the 20:80 combination ratio for AM and ACS showed the best associative effects.     

O-酰基化丝氨酸的合成及其对樱桃番茄乙烯合成量和果实硬度的影响

通过L-丝氨酸与不同的酰氯在室温下反应,合成了8个未见文献报道的乙烯合成抑制剂O-酰基化丝氨酸（2a~2h）,其结构通过核磁共振氢谱、碳谱及高分辨质谱确证。以氨乙氧乙烯基甘氨酸（aminoethoxyvinylglycine,AVG）为阳性对照,测定了目标化合物对樱桃番茄Lycopersivon esculentum Mill.乙烯合成量和果实硬度的影响,同时运用分子对接的方法分析了目标化合物与番茄1-氨基环丙烷羧酸（1-aminocyclopropane-1-carboxylic acid, ACC）合成酶（ACS, 1IAY）可能的结合模式。结果显示:大部分目标化合物具有延缓樱桃番茄果实软化和抑制乙烯合成的作用,其中化合物2c和2h效果较为突出。用2c和2h处理番茄后第5天,与空白对照相比,果实硬度分别提高27.62%和40.04%,均与AVG处理无显著性差异;2c和2h对乙烯合成量的抑制率分别达71.89%和53.28%,其中2c处理的抑制效果显著优于AVG处理。分子对接方法分析结果表明:化合物2c与番茄ACS酶活性空腔的氨基酸残基有较好的相互作用,2c的羧基可与Ala54和Arg412的氨基形成氢键,从而占据ACS酶的活性空腔。O-酰基化丝氨酸类化合物结构简单,容易获取,对乙烯合成抑制剂的开发具有推动作用。     

Role of the genes Md-ACO1 and Md-ACS1 in ethylene production and shelf life of apple (Malus domestica Borkh)

Shelf life determines the economic life time of mature apples, which can be either freshly harvested or stored. Good shelf life is highly associated with a slow decrease of fruit firmness at room temperature. Apple is a climacteric fruit, in which loss of firmness seems to be physiologically related to ethylene. Ethylenes biosynthetic pathway is controlled by two large gene families coding for 1-aminocyclopropane-1-carboxylate synthase (ACS) and 1-aminocyclopropane-1-carboxylate oxydase (ACO).In this study, one ACS and one ACO gene were examined for their effect on ethylene production and shelf life in apple using gene specific molecular marker, and have also been positioned on a molecular marker linkage map. The ACO marker was developed in this research and mapped on linkage group (LG) 10 of the crosses Prima × Fiesta and Fuji × Mondial Gala, within the 5% border of a previously identified fruit firmness QTL [Theor Appl Genet 100 (2000) 1074]. We denoted this locus as Md-ACO1. In addition, we mapped the previously developed Md-ACS1 marker [Theor Appl Genet 101 (2000) 742] on LG15.Studies on the cross Fuji × Braeburn revealed that Md-ACS1 and Md-ACO1 independently affect the internal ethylene concentration (IEC) as well as shelf life of apple, Md-ACS1 having the strongest effect. Descendants homozygous for Md-ACS1-2 and Md-ACO1-1 showed to have the lowest ethylene production as well as superior shelf-life. These two genes are candidates to be included in marker assisted breeding.