Phylogenetic relationships between purple and tea-green individuals of the blue swimming crab Portunus trituberculatus by PCR-SSCP analysis

Given the morphological characters of purple crabs identical with that of the tea-green ones except the body color, it is expected that the purple crabs and the tea-green ones were from the same species. In order to identify the phylogenetic relationships between them at the molecular level, we amplified and compared two conserved mitochondrial gene fragments of purple and tea-green crabs through PCR-SSCP and DNA sequencing analyses. The results showed that there was no distinct variation of DNA bands of both the CO1 and 16srRNA genes detected in polyacrylamide gels through SSCP analysis between purple and tea-green crab samples, and the following sequence analysis of color-different crab individuals presented 99.81% and 99.91% nucleotide sequence identity of the CO1 and 16srRNA genes respectively. These indicated that there was no species or subspecies differentiation between purple and tea-green crabs, meaning that they belonged to the same species, Portunus trituberculatus.     

The Finding and Phylogenetic Evolution Analysis of Bovine Piroplasms in the Rasǒn Area of North Korea

The objective of this study was to investigate the epidemiology of bovine Piroplasms infections in the Rasǒn area of North Korea.The survey was carried out by light microscopic examination of Giemsa-stained blood smears,PCR,and phylogenetic evolution analysis of 128 blood samples collected from the Rasǒn area.The results showed that the infection rates of the small and large parasites were about 2.5 and 1.5% on average,respectively,in all Theileria sergenti and Babesia ovatapositive blood smears by microscopic examination of blood smears.The detection rate of T.sergenti Giemsa-stained smears was 43.75%,while that with PCR was 67.97%.The detection rate of B.ovata Giemsa-stained smears was 49.21%,while that with PCR was 71.88%.The sequence and phylogenetic analysis of DNA showed 98.84% homology between the 18S rRNA gene sequences of T.sergenti isolates from North Korean and that of Yanbian state from China,indicating the closest genetic relationship between both of them.Moreover,100% homology was shown between the 18S rRNA gene sequence of B.ovata isolates from North Korea and the published sequence AY081192 of GenBank,indicating the closest genetic relationship between both of them.This survey confirmed that Ras n is the endemic area of T.sergenti and B.ovata in North Korea.     

Early Stage above Ground Interaction in Wheat-Jatropha Agrisilviculture System

Experiments were conducted to analyze the initial tree crop interaction between Jatropha curcas and wheat by planting the perennial component in East-West orientation with 8-2 m distance and wheat as intercrop at National Research Centre for Agroforestry （NRCAF）, Jhansi during July, 2007 to April, 2009. During planting of Jatropha the interaction to wheat was non-significant and influence of orientation is also nil. Eighteen months after plating Jatropha had attained the mean height of 124.56 cm, collar diameter of 6.37 cm, number of branches of 11.25, canopy diameter of 98.5 cm and yielded 18.5 g seeds per plant. During 18 MAP also observations revealed the non-significant influence of Jatropha on wheat. Observations also indicated the progress in wheat growth and yield attributes with the increase of distances from tree base and it follow the order of 0.5 m〈1.0 m〈2.0 m〈3.0, 4.0 m and open. Nearly, 62% and 14% yield loss in wheat grain was observed at 0.5 m and lm distance from the tree base respectively compare with open. Comparison ofwheat-Jatropha agrisilviculture system and sole wheat cropping indicated the net grain yield reduction up to 13.5% by agrisilviculture system. The comparison of wheat and straw yields between the two years clearly indicating that the system and sole cropping have not much differed while the influence of climatic factor is witnessed by the variation of yields over years. It revealed that the initial above ground interaction between Jatropha and wheat was due to the space that can be managed by the pruning of branches which is a recommended cultural practice of Jatropha.     

Analysis on Genetic Relationship of Oxya chinensis and Oxya japonica from Xuzhou and Pingshan, China

Genetic relationship among Oxya chinensis populations and Oxya japonica populations collected from Xuzhou City, Jiangsu Province and Pingshan County, Hebei Province, China were analyzed by random amplified polymorphic DNA （RAPD） markers. A total of 125 DNA bands ranging from 200 to 2 200 bp were amplified by 10 random primers in DNA samples from 43 grasshopper individuals. One hundred and twenty-three （99%） of these bands were polymorphic. Shannon＇s index showed that the genetic, diversity within O. chinensis （0.3432） was higher than that of O. japonica （0.2781）. Nei＇s genetic distance between O. chinensis population and O. japonica population from the same area was less than that between populations from two different areas. The dendrogram based on Nei＇s genetic distance of RAPD markers was constructed using the unweighted pair group method with the arithmetic average （UPGMA） and Neighbor-Joining （NJ） methods. Cluster analysis indicated that all the individuals were grouped into two main clusters. O. chinensis populations from Xuzhou and Pingshan formed one cluster, and O. japonica populations from the two regions belonged to another cluster. The results demonstrated that RAPD can detect within species, and among closely related species. polymorphisms to distinguish minor difference among individuals     

中国农业科学

In this experiment, the bovine follicular oocytes were aspirated from the ovaries of Chinese Holsteins with laparoscope made in China. The results were as following: for identifying the suitable negative aspiration pressure, six different pressures (50, 100, 150, 200, 250 and 300mmHg)were tested. The aspiration pressure of 100mmHg was the best. Its oocyte recovery rate was 37. 2%, and G I , G Ⅱ oocyte rate was 89. 5%. The heifers were picked up by laparoscope once or twice a week. Each heifer was collected with 2. 4 oocytes once a week or 4. 4 oocytes twice a week.Its oocyte recovery rate was 48. 0% and the G Ⅰ ,G Ⅱ oocyte rate was 93. 5%. In addition, 1.9 oocytes were collected from each cow once a week or 5.4 oocytes from each cow twice a week. Its oocyte recovery rate was 51.7% and the G Ⅰ , G Ⅱ oocyte rate was 85. 1%. It showed that it was possible to pick up bovine oocyte twice a week. Two cows were picked up twice a week for several weeks(53 times). 268 follicles were aspirated(5.1 follicles per cow per time), and 141 oocytes were recovered(2.7 oocytes per cow per time). The oocyte recovery rate was 52.5%, and the G Ⅰ , G Ⅱ oocyte rate was 85. 1%. It was advisable to pick up oocytes twice a week continuously. Some cows in estrous cycles were superovulated with PMSG(500IU). Each of them could be recovered 2.3 follicles and 1.1 oocytes, the others were superovulated with FSH(0. 7mg) , each of them could be aspirated with 4.4 follicles and 2.3 oocytes. It was obvious that the effect of OPU(oocyte pick up) by superovulation with FSH was much better than that with PMSG. The best time for OPU with laparoscope was at the beginning of cow's estrous cycles. At the first day of their estrus, each of them could be averagely aspirated with 8 follicles and 5.7 oocytes.     

苯达松敏感基因在杂交水稻制种技术上的应用研究（英文）

The seed production of male and female parent of 'Hunzhi 1' contained bentazon susceptible gene was studied.The result showed that spraying 3.375 L/hm2 48% bentazon aqua could control the self-pollinated fertilization rate below 1.0% and survival plant rate was 0.The spray of bentazon would not influence MC526 pollen vitality.Seed setting rate of female parent was not obvious different from that of control group with spraying bentazon aqua more than 3.375 L/hm2 from the sprouting appearing period to full heading stage.This demonstrated that bentazon aqua had no influence on seed setting rate during florescence of female parent.The weight ratio between male parent seeds and female parent seeds was 1 to 5,then seeds from parents with that weight ratio would be mixedly planted for seed production.With this weight ratio,relative proper structure of panicle and grain would be obtained,panicle ratio was 1 to 4.3,glumous flowers ratio was 1 to 3.5.Cross seed-setting rate of female parent was 72.6% and seed production per hm2 was 66 000 kg.     

Screening and characterization of a novel ruminal cellulase gene(Umcel-1) from a metagenomic library of gayal(Bos frontalis)

Gayal is a rare semi-wild bovine species found in the Indo-China.They can graze grasses,including bamboo leaves,as well as reeds and other plant species,and grow to higher mature live weights than Yunnan Yellow cattle maintained in similar harsh environments.The aim of this study was to identify specific cellulase in the gayal rumen.A metagenomic fosmid library was constructed using genomic DNA isolated from the ruminal contents of four adult gayals.This library contained38400 clones with an average insert size of 35.5 kb.The Umcel-1 gene was isolated from this library.Investigation of the cellulase activity of 24 random clones led to the identification of the Umcel-1 gene,which exhibited the most potent cellulase activity.Sequencing the Umcel-1 gene revealed that it contained an open reading frame of 942 base pairs that encoded a product of 313 amino acids.The putative gene Umcel-1 product belonged to the glycosyl hydrolase family 5 and showed the highest homology to the cellulase（GenBank accession no.YP₀04310852.1）from Clostridium lentocellum DSM 5427,with 44%identity and 62%similarity.The Umcel-1 gene was heterologously expressed in Escherichia coli BL21,and recombinant Umcel-1 was purified.The activity of purified recombinant Umcel-1 was assessed,and the results revealed that it hydrolyzed carboxymethyl cellulose with optimal activity at pH 5.5 and 45℃.To our knowledge,this study provides the first evidence for a cellulase produced by bacteria in gayal rumen.     

Inheritance of Powdery Mildew Resistance in Cucumber （Cucumis sativus L.） and Development of an AFLP Marker for Resistance Detection

Cucumber powdery mildew is one of the most destructive diseases of cucumber throughout the world. In the present study, inheritance of powdery mildew resistance in three crosses, and linkage of resistance with amplified fragment length polymorphism （AFLP） markers are studied to formulate efficient strategies for breeding cultivars resistant to powdery mildew. The joint analysis of multiple generations and AFLP technique has been applied in this study. The best model is the one with two major genes, additive, dominant, and epistatic effects, plus polygenes with additive, dominant, and epistatic effects （E-l-0 model）. The heritabilities of the major genes varied from 64.26% to 97.82%, and susceptibility was incompletely dominant for the two major genes in the three crosses studied. The additive effects of the two major genes and the dominant effect of the second major gene were high, and the epistatic effect of the additive-dominant between the two major genes was the highest in cross I . In cross II, the absolute value of the additive effect, dominant effect, and potential ratio of the first major gene were far higher than those of the second major gene, and the epistatic effect of the additive-additive was the highest. The genetic parameters of the two major genes in cross III were similar to those in cross II. Correlation and regression analyses showed that marker E25/M63-103 was linked to a susceptible gene controlling powdery mildew resistance. The marker could account for 19.98% of the phenotypic variation. When the marker was tested on a diverse set of 29 cucumber lines, the correlation between phenotype and genotype was not significant, which suggested cultivar specialty of gene expression or different methods of resistance to powdery mildew. The target DNA fragment was 103 bp in length, and only a small part was found to be homologous to DNA in the other species evaluated, which indicated that it was unique to the cucumber genome.     

Studies on the Host Biotypes and Its Cause of Cotton Aphid in Nanjing, China

The fitness of cotton aphid, Aphis gossypii Glover, on different host plants was studied. The results showed that the cotton aphid population in Nanjing formed host preference bio-types on cucumber and cotton. The aphids from cotton and cucumber could not inter-transplant successfully between the two host plants. The apterous and alate offspring of stem mother from hibiscus could not survive, reproduce and form population normally when they were transplanted to cucumber and water melon, but they could form normally when transplanted to cotton and muskmelon. There was no significant difference between the stylet length of cotton aphids from cucumber and cotton, so the stylet length was not the cause of the host preference. When the aphids originated on hibiscus were transplanted to cotton, water melon and muskmelon, the stylet could reach host phloem to absorb host plant‘s nutrients, but it couldn‘t when transplanted to cucumber. Cotton aphid can feed on muskmelon without secretion of water-soluble saliva. It could be concluded that the two host biotypes of cotton aphid came from different winter host plants.