南荻同源四倍体的研究

南荻变种“突节荻”的幼穗、下胚轴产生的带绿芽点愈伤组织,置于０．０５％秋水仙碱的ＭＳ无激素培养基上处理２４－４８ｈ,经培养而诱变成交异试管苗。细胞学鉴定确定其染色体数目为２ｎ＝４ｘ＝７６（ｘ＝１９）,花粉粒和气孔保卫细胞均增大,核仁数目增多。田间种植和测定,获得南荻同源四倍体新品系。该品系表现出生长势强,植株高大,茎粗、茎壁厚、节间长,产量高,而且纤维长,长宽比值大,是生产上能直接利用的一种优质高产的造纸新资源     

两种农业种植模式对重金属土壤的修复潜力

植物修复农田土壤重金属污染需要经历一个长期的过程,而大部分用来修复的植物都不具备利用价值,不能给当地农民带来经济收入。因此,一些农作物由于其较大的生物量和一定的经济价值,在植物修复土壤重金属污染的应用中受到广泛关注。是在重金属(Cu、Zn、Pb、Cd、As、Hg)复合污染的郴州矿区废弃农田种植油菜、玉米和油葵,研究油菜-玉米和油-油葵两种种植模式对土壤重金属污染的修复潜力。实验结果表明:三种作物在复合污染土壤中对重金属都表现出一定的耐性及吸收积累能力。向日葵的根和叶中重金属Cd、Cu的含量都很高,其中Cd在向日葵的各个部位的富集系数(BCF)及Cu在向日葵的根和叶的富集系数(BCF)都大于1。两种轮作模式对作物的产量没有明显的影响,收获得到的干物质量都很高,每年每公顷分别为油菜16.6t、玉米25.29t、油葵22.5t。两种种植模式都可以对土壤中的重金属进行有效的提取,油菜-油葵种植模式下提取重金属Cu、Pb、Cd、As的量较高,分别为:Cu 2408g hm~(-2)a~(-1),Pb 2027g hm~(-2)a~(-1),Cd 658.5g hm~(-2)a~(-1),As 250g hm~(-2)a~(-1),油菜-玉米模式下Zn和Hg的提取量较高,分别为Zn 4987g hm~(-2)a~(-1)和Hg 7.92g hm~(-2)a~(-1);对于多种重金属复合污染的土壤来说,油菜-油葵的种植模式要优于油菜-玉米的种植模式。总的来说,利用3种作物两两轮作的种植模式,在不影响作物产量的前提下大大的提高了作物对重金属的提取总量。3种作物在收获以后又可以用做工业原料,这就使得当地农民充分利用矿区废弃农田修复污染的同时又能从中获得一定经济效益。     

花期渍水逆境下氮素对油菜产量及氮肥利用效率的影响

温室盆栽试验条件下,设置渍水和对照2个水分处理,每个水分处理下设置3个施氮水平(0.05、0.2、0.3 g N·kg^-1土),研究了花后渍水逆境下氮素营养对两个氮高效基因型‘Monty’、‘湘油15’和两个氮低效基因型‘R210’、‘Bin270’油菜产量、产量性能及氮肥利用效率的影响.结果表明:与对照相比,花后渍水处理显著降低了油菜的单株角果数、千粒重、每角粒数和籽粒产量.在适宜水分条件下,增施氮肥显著增加了油菜籽粒产量,而在渍水逆境处理下,增施氮肥对油菜籽粒产量的形成贡献不大.氮高效基因型较氮低效基因型对花后渍水逆境下的籽粒灌浆充实具有一定的促进作用.在同一水分处理下,花后渍水明显降低了油菜氮肥利用率、氮肥偏生产力、氮肥农学利用率、氮素吸收效率和氮收获指数,渍水显著影响了不同基因型油菜的氮素吸收利用能力,而氮高效基因型在渍水逆境下较氮低效基因型更有利于将氮素转运、再分配到角果中,提高籽粒生产效率.油菜产量性能参数存在显著的水氮互作效应,水分、氮肥及水氮互作对油菜籽粒产量和产量性能参数的影响因基因型的不同而异.     

The role of plants in the removal of nutrients at a constructed wetland treating agricultural (dairy) wastewater,Ontario, Canada

The South Nation River Watershed, in eastern Ontario, Canada, is an agricultural watershed impacted by excess nutrient loading primarily from agricultural activities. A constructed wetland for the treatment of agricultural wastewater from a 150-cow dairy operation in this watershed was monitored in its eighth operating season to evaluate the proportion of total nitrogen (TN) (approximated by total Kjeldahl nitrogen (TKN) due to low NO₃⁻) and total phosphorus (TP) removal that could be attributed to storage in Typha latifolia L. and Typha angustifolia L., which dominate this system. Nutrient loading rates were high, with 16.2 kg ha⁻¹ d⁻¹ N and 3.4 kg ha⁻¹ d⁻¹ P entering the wetland and loading the first wetland cell. Plant uptake accounted for 0.7% of TKN removal when the vegetated free water surface cells were considered together. However, separately, in the second wetland cell with lower N and P loading rates, plants accounted for 9% of TKN, 21% of NH₄⁺ and 5% of TP removal. Plant uptake was significant to overall removal given wetland age and nutrient loading. Nutrient storage during the growing season at this constructed wetland helped reduce the nutrient load entering the watershed, already stressed by intensive local agriculture.     

枯草杆菌manA基因的克隆及定点突变

manA是编码β-甘露聚糖酶(β-1,4-mannan mannohydrolase EC3.2.1.78)的基因。将枯草杆菌A33株的manA基因插入到pET-32a载体,并在大肠杆菌BL21(DE3)中实现了异源非融合表达,表达活力为41.58U/mL。为了提高酶的表达活力,当采用PCR介导的定点突变技术将该基因第2号密码子CUU突变为GUU,构建成突变表达载体pET-32a-manA*并转入大肠杆菌BL21(DE3)中表达,目标酶表达活力增加到138.65U/mL。说明当β-甘露聚糖酶N端第二号氨基酸由亮氨酸突变为缬氨酸后,酶在大肠杆菌中的表达活力大大提高。推测是由于突变后的β-甘露聚糖酶在大肠杆菌中的稳定性增强所致。突变表达的β-甘露聚糖酶最适作用温度和pH值并没有发生明显改变。     

Role of iron plaque in Cd uptake by and translocation within rice (Oryza sativa L.) seedlings grown in solution culture

A hydroponics culture experiment was conducted to investigate the effect of iron plaque on Cd uptake by and translocation within rice seedlings grown under controlled growth chamber conditions. Rice seedlings were pre-cultivated for 43 days and then transferred to nutrient solution containing six levels of Fe (0, 10, 30, 50, 80 and 100 mg L⁻¹) for 6 days to induce different amounts of iron plaque on the root surfaces. Seedlings were then exposed to solution containing three levels of Cd (0, 0.1 and 1.0 mg L⁻¹) for 4 days. In order to differentiate the uptake capability of Cd by roots with or without iron plaque, root tips (white root part without iron plaque) and middle root parts (with iron plaque) of pre-cultivated seedlings treated with 0, 30 and 50 mg L⁻¹ Fe were exposed to ¹⁰⁹Cd for 24 h. Reddish iron plaque gradually became visible on the surface of rice roots but the visual symptoms of the iron plaque on the roots differed among treatments. In general, the reddish color of the iron plaque became darker with increasing Fe supply, and the iron plaque was more homogeneously distributed all along the roots. The Fe concentrations increased significantly with increasing Fe supply regardless of Cd additions. The Cd concentrations in dithionite–citrate–bicarbonate (DCB)-extracts and in shoots and roots were significantly affected by Cd and Fe supply in the nutrient solution. The Cd concentrations increased significantly with increasing Cd supply in the solution and were undetectable when no Cd was added. The Cd concentrations in DCB-extracts with Fe supplied tended to be higher than that at Fe₀ at Cd_0.1, and at Cd_1.0, DCB-Cd with Fe supplied was significantly lower. Cd concentrations in roots and shoots decreased with increasing Fe supply at both Cd additions. The proportion of Cd in DCB-extracts was significantly lower than in roots or shoots. Compared to the control seedlings without Fe supply, the radioactivity of ¹⁰⁹Cd in shoots of seedlings treated with Fe decreased when root tips were exposed to ¹⁰⁹Cd and did not change significantly when middle parts of roots were exposed. Our results suggest that root tissue rather than iron plaque on the root surface is a barrier to Cd uptake and translocation within rice plants, and the uptake and translocation of Cd appear to be related to Fe nutritional levels in the plants.     

高效的'大红'甜橙实生苗上胚轴离体培养与植株再生体系的建立

以‘大红’甜橙实生苗上胚轴为外植体,研究其离体培养和植株再生技术的结果表明,MS 1.0mg·L-16-BA的培养基诱导不定芽的效果最好,30d时诱导率达97.5%。外植体抗性筛选出最适kanamycin浓度为50mg·L-1。IBA和NAA都促进不定芽生根。1/2MS 4.0mg·L-1IBA和1/2MS 1.5mg·L-1NAA诱导根的效果最佳,生根率和根数分别达到76.92%、0.81和92.31%、1.88。     

长期施肥对茶园土壤微生物群落功能多样性的影响

利用BIOLOG研究长期施肥后茶园土壤微生物群落功能多样性变化,结果表明:与清耕(CK)相比,各长期施肥处理AWCD值的变化速度(斜率)和最终能达到的AWCD值均有不同程度提高,其大小顺序为:DWY>DY>W>CK,可见施肥能不同程度提高微生物整体活性和丰富度,以有机无机肥配施效果最好。分析多样性指数表明:各处理对土壤常见微生物种类影响并不大,施有机肥使微生物群落均匀度有所降低。对碳源利用主成分起分异作用的主要是糖类和羧酸类物质。对AWCD值、多样性指数与土壤养分因子的相关性分析表明,在红壤茶园中土壤肥力提高有利于土壤微生物活性的提高。     

与烟草细胞质雄性不育相关的线粒体基因atp9的mRNA研究

已知导入未编辑atp9 mRNA的烟草表现细胞质雄性不育(CMS),因此认为线粒体基因atp9是引起高等植物CMS的主要基因。为了解atp9在CMS中的作用机制,从3对烟草不育系及其同型保持系中提取atp9,利用实验与理论结合来分析其mRNA在编辑前后以及在不育系及其同型保持系中的一维、三维信息差别。结果表明,atp9mRNA一维信息方面的差异,更重要的是二级结构的差异和稳定性,可能是影响ATP合成而导致CMS的根本原因。     

Enhancement of edeine production in Brevibacillus brevis X23 via in situ promoter engineering

Edeines, a group of cationic antimicrobial peptides produced by the soil bacterium Brevibacillus, have broad biological effects, such as antimicrobial, anticancer and immunosuppressive activities. However, the yield of edeines in wild-type (WT) Brevibacillus is extremely low, and chemical synthesis of edeines is a time-consuming process. Genetic engineering has proven to be an effective approach to produce antibiotics with high yield. In this study, the ede ine b iosynthetic g ene c luster (ede BGC), which is involved in edeine production, was identified and characterized in Brevibacillus brevis X23. To improve edeine production in B. brevis X23, the ede BGC promoter was replaced with six different promoters, P_mwp, P_spc, P_xylA, P_shuttle-09, P_grac or P₄₃, through double-crossover homologous recombination. The new promoters significantly increased the expression of the ede BGC as well as edeine production by 2.9 ± 0.4 to 20.5 ± 1.2-fold and 3.6 ± 0.1to 8.7 ± 0.7-fold respectively. The highest yield of edeines (83.6 mg l⁻¹) was obtained in B. brevis X23 with the P_mwp promoter. This study provides a practical approach for producing high yields of edeines in B. brevis.