Simple method of genomic DNA extraction from Rhizobia in dried nodules of Phaseolus vulgaris for strain differentiation by PCR-based DNA fingerprinting

Repetitive DNA peR fingerprinting of bacterial genomic DNA is a useful tool for typing and differentiation of rhizobial strains. The method was reported to be suitable for strain differentiation of Rhizobia present in individual root nodules of some leguminous plants without the need for isolation and cultivation of the strains, in which rhizobial genomic DNA was extracted directly from each fresh or frozen nodule. We developed a new protocol of rhizobial genomic DNA extraction/purification from dried nodules of Phaseolus vulgaris for generating repetitive DNA peR fingerprints of Rhizobia present in the nodules. The simplified protocol consists of only three major steps, heat extraction of genomic DNA from rhizobial cells prepared from dried nodules, ethanol precipitation of the DNA and Sephadex G-50 column purification of the DNA, and generated fingerprints with good quality for differentiation of Rhizobia strains. The protocol will be useful to examine the nodule occupancy of inoculated rhizobial strains in field experiments.     

酿酒酵母耐铜基因CUP1的克隆及其植物表达载体的构建

酿酒酵母铜金属硫蛋白（ｃｏｐｐｅｒ ｍｅｔａｌｌｏｔｈｉｏｎｅｉｎ,Ｃｕ－ＭＴ）,属于第Ⅱ型金属硫蛋白,Ｃｕ－ＭＴ由酿酒酵母耐铜基因ＣＵＰ１编码,由６１个氨基酸残基组成,其中半胱氨酸残基１３个,占Ｃｕ－ＭＴ总氨基酸数的２１．３％,Ｃｕ－ＭＴ与铜离子有较强的结合力,在酵母细胞中主要参与过量铜的解毒作用。本研究通过ＰＣＲ方法,从酵母基因组ＤＮＡ中扩增出了ＣＵＰ１基因,克隆于ｐＵＣ１９的多克隆拉点。扩增     

镉胁迫下大豆中镉与几种微量元素的分布状况

镉是一种毒性较大的重金属元素,易在作物中积累,通过食物链危害动物和人类健康。日本发生的骨痛病便是镉污染的典型病症。我国有1万多公顷农田遭受镉污染,镉污染直接导致农产品品质恶化,在一些地区已出现了镉中毒人群。 　　大豆是植物性蛋白和油脂的主要来源,也是重要的食粮和精饲料,我国是大豆的主产国也是最大的消费国。由于大豆具有较强的耐旱、耐贫脊的特点,往往作为废矿复垦等污染区的主要作物。因此,弄清镉在大豆中（油分、粗蛋白和淀粉）的分布状况对于合理加工利用污染区农产品、切断重金属在食物链中的传递具有一定环保和经济意义。本研究初步探明了镉胁迫下大豆对镉与微量元素营养元素吸收,及其在大豆植株、籽粒、油分和豆粕中的分布状况,为利用基因工程改良大豆的重金属耐性积累了基础资料...     

Treponemes-Infected Canker in a Japanese Racehorse: Efficacy of Maggot Debridement Therapy

A 3-year-old thoroughbred colt presented with canker on its left hind foot. Subsequent development of cottage cheese-like horns and dermatitis disturbed healing, despite the use of miscellaneous orthodox treatment approaches to the lesions. Histological examination revealed exudative and suppurative dermatitis, and proliferatively suppurative epidermitis infected with helically coiled treponemes. Total debridement under general anesthesia led to a temporary improvement, but the ground surface regenerated abnormal epidermis similar to that observed initially after surgery. Maggot debridement therapy (MDT) was attempted, which removed all the abnormal tissue. After MDT, general farriery trimming helped to correct the distorted ground surface, and the horse returned to constant training and eventually raced. This case shows that MDT was successfully used for treatment of an intractable and treponemes-infected canker.     

Decontamination of Chlorate in Longan Plantation Soils by Bio-Stimulation with Molasses Amendment

Potassium chlorate is widely used as an active substance for flower induction in longan plantation fields for off-season production of longan fruits in northern Thailand. Contamination of groundwater with residual chlorate in soil is a cause for concern because of its toxicity to human health. Based on our previous finding that the addition of glucose or sucrose to soil was effective for accelerating the disappearance of residual chlorate in soil, the effect of the addition of molasses, which contains a high concentration of sucrose, as a substitute for glucose or sucrose was examined in laboratory and pot experiments. Under laboratory conditions, the addition of molasses to soil at the concentrations of 100 to 200 g kg⁻¹ soil strikingly enhanced the rate of disappearance of chlorate applied at 341 mg kg⁻¹ soil. Addition of diluted molasses was also effective for the accelerated disappearance of chlorate in soil when 33 g kg⁻¹ soil of molasses was added repeatedly. The effect of repeated addition of diluted molasses to soil on the decontamination of residual chlorate in soil was also confirmed in an outdoor pot experiment. These results may lead to the development of a practical method of cleaning-up chlorate-polluted soil in longan plantation fields.     

Tracer experiments on transfer of radio-iodine in the soil — rice plant system

In order to study the transfer of ¹²⁹I from soil into rice, tracer experiments have been carried out in the soil-rice plant system. The soil-plant transfer factors, which are defined as ‘concentrations of the nuclide in a plant organ’ divided by ‘concentrations of the nuclide in dry soil’, were 6.0 × 10^?3 for brown rice, 3.2 for the first leaf blade (flag leaf), 5.1 for the second leaf blade, 2.6 for stem, 1.4 for rachis and 9.0 for root on a dry weight basis at the time of harvest. The ratio of radio-iodine in brown rice to polished rice was about 1:0.3. The transfer factor of radio-iodine from soil to polished rice was estimated to be about 2 × 10^?3. It was observed that the concentration of radio-iodine in the soil solution increased with the plant growth. This was attributed to the reduced condition in the soil caused by the effects of root and/or micro-organisms. Release of I from rice plants into the air in an organic form was found. The chemical form of radio-iodine in the soil solution appeared to be iodide, and the radio-iodine in the plant tissue was found in a low molecular weight fraction by gel-filtration. The vertical distribution of radioiodine in the soil was also studied.     

Intestinal spirochetosis in a 21-month-old thoroughbred colt

A 21-month-old Thoroughbred colt showed continuous diarrhea and developmental retardation for 7 months, and was thereafter subjected to euthanasia for necropsy and laboratory examinations. At necropsy, the cecal and colonic mucosae were diffusely rough and hyperemic. Histopathologically, the mucosa and submucosa were edematous and were infiltrated by numerous lymphocytes and macrophages. Meanwhile, three morphological types of Brachyspira antigen-containing spirochetes were found to be numerous in the crypts and in the mucus layer over the epithelium in the cecal and colonic lesions. They were frequently observed in intercellular gaps and in the cytoplasm of degenerative epithelial cells, and in the lamina propria, particularly in cavities around blood vessels. These invasive intestinal spirochetes might be one of pathogens inducing colitis and diarrhea in horses.