The Suaeda liaotungensis kitag betaine aldehyde dehydrogenase gene improves salt tolerance of transgenic maize mediated with minimum linear length of DNA fragment

In this research we established a particular vector-free and marker-free plant transformation system of maize to overcome the obstacles of biosafety limits. The BADH gene was introduced into maize by pollen-tube pathway, using the principle of minimum linear length of the transformation element, which was composed of only the BADH gene, expression regulatory sequence (35S CAMV promoter, NOS terminator), and T-DNA border sequence at both sides. Twenty-seven of 2076 transformed samples were positive in PCR amplification and the PCR positive rate of T₁ generation was 1.3%. Further Southern blotting results indicated that the BADH gene was integrated into maize genome. Transgenic lines of progeny were examined for tolerance to NaCl by induced salt stress with 250 mM NaCl Hoagland solution. After 15 days of treatment, 73.9–100% of the transgenic seedlings survived and grew well, whereas most wild-type seedlings wilted and showed loss of chlorophyll. Only 8.9% of the wild-type plants survived but gradually died after salt stress. The electrical conductivity of the transgenic line of progeny after salt stress was lower than wild type. The transgenic progeny had higher glycinebetaine and Chlorophyll content than wild type after salt stress.     

2,3-丁二醇脱氢酶基因(BudC)过表达菌株的构建和初步鉴定

旨在过表达BudC进而上调2,3-丁二醇（2,3-Butanediol, 2,3-BD）的合成,通过基因工程手段构建整合载体pR1SW-BudC,经PCR和酶切双重验证后将其电转化至产酸克雷伯氏菌(Klebsiella oxytoca) HD79。结果经卡那霉素筛选,获得一株菌株K. oxytoca HD79-01。以原始菌株为对照,摇瓶发酵检测2,3-BD含量和相关酶活,q RT-PCR检测BudC表达差异,最终BudC在HD79-01中转录水平的表达量为HD79的45.25倍（p<0.01）。摇瓶发酵显示2,3-BD的最高产量、转化率和生产强度分别提高了24.54%、10.97%、45.08%（分别为30.818 ± 0.003 g/L、0.253 ± 0.013 g/g、0.43 ± 0.01 g/(L·h)）,酶活提高了3.61倍(0.563 ± 0.003 U/mg)。因此,2,3-丁二醇脱氢酶基因(BudC)过表达菌株构建不仅成功的提高了2,3-BD的产量也为实现2,3-BD的工业化生产奠定基础。     

转甜菜碱醛脱氢酶基因水稻种子萌发和苗期耐盐性的研究

用0.0、3.0、5.0g/L NaCl对2个转甜菜碱醛脱氢酶(BADH)基因水稻51-22、52-7与受体亲本中花8进行萌发期盐胁迫试验.结果表明,盐胁迫下转基因材料受抑程度小于受体亲本,尤以52-7表现明显.用0.0、3.0、5.0g/L NaCl对51-22、52-7和中花8在幼苗期进行14 d盐胁迫试验.结果表明,低盐胁迫时幼苗受盐害较小,高盐胁迫时转基因材料耐盐性优于受体亲本;在盐胁迫7 d后,转基因材料体内脯氨酸含量明显高于受体亲本,同一品种随盐度增大,脯氨酸含量升高.     

Characterization of biochar and their influence on microbial activities and potassium availability in an acid soil

Application of biochar to soil has increased considerably during recent years because of its effectiveness as a soil amendment causing beneficial effects on soil health. However, the effects have been reported to vary and depend upon types of feedstock and pyrolysis conditions during biochar production. Therefore, characterization of biochar is extremely important for its efficient utilization as a soil amendment. In the present study, biochar was prepared from agro-industrial by-products (rice husk and sugarcane bagasse) and weeds (Parthenium and Lantana) under similar pyrolysis conditions. Lantana biochar (LBC) showed the highest pH (10.4) while the lowest value (8.5) being recorded in rice husk biochar (RHBC). The energy-dispersive X-ray spectroscopy (EDS) analysis indicated that LBC and Parthenium biochar (PBC) were superior with respect to potassium (K) content than sugarcane bagasse biochar (SBBC) and RHBC. The Fourier-Transform Infrared Spectroscopy (FTIR) study exhibited the existence of different functional groups in biochar. All the biochar treated soils showed significantly higher microbial activities with different degrees. Application of LBC and PBC at 4.50 g kg^?1 soil significantly increased K availability in soil. Lantana biochar and PBC amended the soil at 9 g kg^?1 significantly increased the soil pH thus makes these biochar as potential liming materials.     

转甜菜碱醛脱氢酶基因马铃薯的抗旱耐盐性

通过根癌农杆菌介导法将甜菜碱醛脱氢酶(BADH)基因导入马铃薯栽培品种甘农薯2号, 经PCR、Southern杂交和Northern杂交证明BADH基因已整合到马铃薯基因组中并在转基因植株中转录和表达。测定表明对照植株没有BADH酶活性, 各转化株系在胁迫前后BADH酶活性近似, 在2~11 U之间。BADH酶活性与叶片的相对电导率呈一定的负相关(y= –3.7738x+57.083, r=0.989^**)。在NaCl和PEG胁迫下, 转基因植株生长正常, 株高比对照提高0.41~1.00 cm, 单株重量比对照增加10%~35%, 说明外源BADH基因的导入提高了马铃薯植株对干旱和盐碱的抗性。     

转甜菜碱醛脱氢酶基因提高烟草抗旱及耐盐性

将甜菜碱醛脱氢酶（BADH）基因与组成型启动子CaMV 35S启动子融合,构建了植物表达质粒pBIBB。通过根癌农杆菌介导将BADH基因导入烟草,经PCR、Southern杂交、Northern杂交证明BADH基因已整合到烟草基因组中并在转基因植株中转录和表达。测定转基因植株叶片中甜菜碱醛脱氢酶活性,结果显示对照植株没有BADH酶活性,转基因植株的各个株系间甜菜碱醛脱氢酶比活力差异较大,范围在0.1~1.0 U mg-1间。转BADH基因的烟草在盐胁迫和聚乙二醇（PEG）胁迫条件下生长状态良好,生长势强于未转基因植株,说明BADH基因能在异源植物中正常翻译、表达和用于植物抗旱、耐盐基因工程的研究。     

Generation and analysis of expressed sequence tag sequences from a soft‐seeded pomegranate cDNA library

The cultivation of soft‐seeded pomegranate is an important direction in pomegranate breeding. To comprehensively understand the molecular mechanisms involved in the formation of soft‐seeded pomegranate (Punica granatum. var. Hongmanaozi), we established an expressed sequence tag (EST) resource. Two thousand valid sequences were generated, from which 907 unigenes were obtained after initial assembly using the clustalx program. Among these unigenes, 51 showed no similarity to any protein in the public databases, 433 matched with proteins of unknown function, and 423 matched with proteins of known or putative functions. The 423 unigenes were further classified into 13 categories. Among these categories, protein synthesis, cell structure, protein destination and storage, secondary metabolism, signal transduction and transporters accounted for 8%, 8%, 4%, 7%, 6% and 17%, respectively. We also successfully developed 10 highly polymorphic expressed sequence tag‐simple sequence repeat (EST‐SSR) markers for pomegranate. The results provide a new tool for future activities in pomegranate breeding.     

低温胁迫对番茄幼苗叶片中脯氨酸降解酶的活性及其基因表达量的影响

为了探讨脯氨酸代谢与番茄抗寒性之间的关系,采用常规方法测定了低温胁迫不同时间下番茄幼苗叶片中游离脯氨酸含量和脯氨酸脱氢酶活性,采用实时荧光定量PCR法检测了不同处理时间下脯氨酸脱氢酶基因的表达量,并分析了各项指标的动态变化。结果表明,在低温逆境下,耐寒性品种O-33-1中脯氨酸积累量总体呈增加的趋势,而冷敏感品种‘耐运2000’游离脯氨酸积累量下降,说明脯氨酸积累量的变化在一定程度上可以体现番茄抗寒性的强弱;ProDH活性在O-33-1中呈上升趋势,在‘耐运2000’中呈下降趋势,说明脯氨酸的积累受P5CS与ProDH相反方向的共同调节,使植物体内脯氨酸含量保持在适当的水平;2个品种在低温胁迫下ProDH基因表达量都显著降低,说明低温逆境抑制了该基因的表达,使脯氨酸的降解在转录水平上受到调节。     

不同产地人参中4种脱氢酶活力比较

为了给人参的培育和优选提供理论依据,采用中性缓冲液提取粗酶液,应用分光光度法对15个不同产地的人参中苹果酸脱氢酶(Malate Dehydrogenase MDH),乳酸脱氢酶(Lactate Dehydrogenase, LDH),乙醇脱氢酶(Alcohol Dehydrogenase, ADH),葡萄糖-6-磷酸脱氢酶(Glucose-6-phosphate Dehydrogenase, G6PDH）4种脱氢酶活力进行比较。结果表明不同产地人参脱氢酶活力差别明显,同一产地4种脱氢酶活力趋势基本相同。其中安图县万宝镇的人参样品的MDH、LDH、G6PDH 3种酶活力均是最高值,分别为124.58U/(g?Fw)、129.88 U/(g?Fw)、109.84U/(g?Fw);黑龙江2个产地的4种酶活力普遍比较低。运用SPSS13.0软件对15批样品进行系统聚类分析,将样品分为4类。MDH、LDH、ADH、G6PDH的活力可以作为人参培育和优选提供理论依据。