玉米ZmPTF1基因克隆和过表达分析

PTF是一类具有bHLH结构域的转录因子,在低磷胁迫应答中调节一系列下游磷胁迫基因的表达.根据拟南芥和水稻中的序列信息,通过RACE的方法获得ZmPTF1的全长cDNA序列.序列分析表明该1709bp的cDNA片段含有1446bp的完整开放读码框,编码481个氨基酸的多肽链,含有一个bHLH结构域,与已报道的OsPTF1在氨基酸水平上有65%的相似性.构建了pCAMBIA1300-Pubi-ZmPTF1-Tnos-als质粒转化玉米,得到过表达的转基因后代.对低磷营养液浇灌下的T2代植株分析表明:过表达ZmPTF1促进了植株根系的发育,提高了植株对低磷环境的耐受性.     

Overexpression of a Cytosolic Ascorbate Peroxidase Gene,OsAPX2, Increases Salt Tolerance in Transgenic Alfalfa

Alfalfa （Medicago sativa L.） is an important forage crop in the world and it is of great signiifcance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene （OsAPX2） was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation with marker gene bar. The different T-DNA insertions in T1 transgenic alfalfa were identiifed by Southern hybridization. Three independent T2 transgenic lines were selected for stress analysis and the results showed that all of them were salt tolerant compared with wild-type plants. The transgenic plants had low levels of H2O2, malondialdehyde and relative electrical conductivity under salt and drought stresses. Moreover, the contents of chlorophyll and proline, and APX activity were high in transgenic plants under salt and drought stresses. Taken together, the overexpression of OsAPX2 enhances salt tolerance in alfalfa through scavenging reactive oxygen species.     

转GmPTF1大豆耐低磷优异新材料创制研究

本试验以课题组前期获得的转耐低磷转录因子GmPTF1大豆为材料,分析其在低磷、适磷条件下的性状表现,研究其在来源物种中的生物学功能,并遴选优异转基因新材料。结果表明:目的基因GmPTF1已稳定整合至受体品种基因组,且可在RNA和蛋白水平正常转录翻译表达;土壤低磷处理条件下,2个转基因株系OE-2、OE-4单株荚数、单株粒数、单株粒重等产量相关性状显著优于野生型对照,并以OE-2株系表现较优,为进一步选育耐低磷大豆新品系(种)奠定了重要物质基础。     

Genome-Wide Association Study for Certain Carcass Traits and Organ Weights in a Large White×Minzhu Intercross Porcine Population

Porcine carcass traits and organ weights have important economic roles in the swine industry. A total of 576 animals from a Large White×Minzhu intercross population were genotyped using the Illumina PorcineSNP60K Beadchip and were phenotyped for 10 traits, speciifcally, backfat thickness （6-7 libs）, carcass length, carcass weight, foot weight, head weight, heart weight, leaf fat weight, liver weight, lung weight and slaughter body weight. The genome-wide association study （GWAS） was assessed by Genome Wide Rapid Association using the mixed model and regression-genomic control approach. A total of 31 single nucleotide polymorphisms （SNPs） （with the most signiifcant SNP being MARC0033464, P value=6.80×10-13） were located in a 9.76-Mb （31.24-41.00 Mb） region on SSC7 and were found to be signiifcantly associated with one or more carcass traits and organ weights. High percentage of phenotypic variance explanation was observed for each trait ranging from 31.21 to 67.42%. Linkage analysis revealed one haplotype block of 495 kb, in which the most signiifcant SNP being MARC0033464 was contained, on SSC7 at complete linkage disequilibrium. Annotation of the pig reference genome suggested 6 genes （GRM4, HMGA1, NUDT3, RPS10, SPDEF and PACSIN1） in this candidate linkage disequilibrium （LD） interval. Functional analysis indicated that the HMGA1 gene presents the prime biological candidate for carcass traits and organ weights in pig, with potential application in breeding programs.     

Transgenic cry1Ab/vip3H＋epsps Rice with Insect and Herbicide Resistance Acted No Adverse Impacts on the Population Growth of a Non-Target Herbivore,the White-Backed Planthopper,Under Laboratory and Field Conditions

Numerous Bt rice lines expressing Cry protein derived from Bacillus thuringiensis Berliner （Bt） have been developed since 1989. However, the potential risks posed by Bt rice on non-target organisms still remain debate. The white-backed planthopper （WBPH）, Sogatella furcifera （Horváth）, is one of the most economically important insect pests of rice in Asian countries and also one of the main non-target herbivores of transgenic rice. In the current study, impacts of transgenic cry1Ab/vip3H＋epsps rice （G6H1） with both insect and herbicide resistance on WBPH were evaluated to ascertain whether this transgenic rice line had potential risks for this sap-sucking pest under laboratory and ifeld conditions. The laboratory results showed that no signiifcant difference in egg developmental duration, nymphal survival rate and female fecundity was found for WBPH between G6H1 and its non-transgenic isoline （XS110）. However, the development duration of nymphs was signiifcantly shorter and female longevity signiifcantly longer when WBPH fed on G6H1 by comparison with those on its control. To verify the results found in laboratory, a 3-yr ifeld trial was conducted to monitor WBPH population using both the vacuum-suction machine and beat plate methods. Although the seasonal density of WBPH nymphs and total density of nymphs and adults were not signiifcantly affected by transgenic rice regardless of the sampling methods, the seasonal density of WBPH adults in transgenic rice plots was slightly lower than that in the control when using the vacuum-suction machine. Based on these results both from laboratory and ifeld, it is clear that our tested transgenic rice line will not lead higher population of WBPH. However, long-term ifeld experiments to monitor the population dynamics of WPBH at large scale need to be conducted to conifrm the present conclusions in future.     

过量表达AtNHXS1新基因显著提高水稻的耐盐性

以转基因和野生型水稻为材料,通过农杆菌介导法将AtNHXS1转到水稻植株中花11号中,分析在盐胁迫下Na+、K+含量的变化,对两者耐盐性进行比较,并对转基因株系进行分子鉴定和转录表达分析。结果表明:PCR初步鉴定得到了20个转基因株系,随机挑选2个PCR阳性株系进行Southern blot鉴定,确定AtNHXS1以单拷贝的形式成功插入到水稻基因组中。耐盐性分析表明,在盐胁迫条件下,转基因水稻植株的生长状况、干质量、鲜质量、Na+含量显著优于或高于野生型水稻植株;此外,300mmol/L NaCl处理下,转基因水稻植株能够正常存活,而野生型水稻5d内几乎全部死亡。将300mmol/L NaCl处理过的植株在无盐胁迫的条件下进行恢复生长试验,转基因植株10d内恢复正常,而野生型则不能。过量表达改组后的AtNHXS1新基因显著提高了水稻的耐盐性。     

Transgenic Expression of a Functional Fragment of Harpin Protein Hpa1 in Wheat Represses English Grain Aphid Infestation

The harpin protein Hpa1 produced by the rice bacterial blight pathogen promotes plant growth and induces plant resistance to pathogens and insect pests. The region of 10-42 residues （Hpa110-42） in the Hpa1 sequence is critical as the isolated Hpa110-42 fragment is 1.3-7.5-fold more effective than the full length in inducing plant growth and resistance. Here we report that transgenic expression of Hpa110-42 in wheat induces resistance to English grain aphid, a dominant species of wheat aphids. Hpa110-42-induced resistance is effective to inhibit the aphid behavior in plant preference at the initial colonization stage and repress aphid performances in the reproduction, nymph growth, and instar development on transgenic plants. The resistance characters are correlated with enhanced expression of defense-regulatory genes （EIN2, PP2-A, and GSL10） and consistent with induced expression of defense response genes （Hel, PDF1.2, PR-1b, and PR-2b）. As a result, aphid infestations are alleviated in transgenic plants. The level of Hpa110-42-induced resistance in regard to repression of aphid infestations is equivalent to the effect of chemical control provided by an insecticide. These results suggested that the defensive role of Hpa110-42 can be integrated into breeding germplasm of the agriculturally signiifcant crop with a great potential of the agricultural application.     

Fruit Yield and Quality,and Irrigation Water Use Efifciency of Summer Squash Drip-Irrigated with Different Irrigation Quantities in a Semi-Arid Agricultural Area

Fruit yield, yield components, fruit mineral content, total phenolic content, antioxidant activity and irrigation water use efifciency （IWUE） of summer squash responses to different irrigation quantities were evaluated with a ifeld study. Irrigations were done when the total evaporated water from a Class A pan was about 30 mm. Different irrigation quantities were adjusted using three different plant-pan coefifcients （Kcp, 100% （Kcp1）, 85% （Kcp2） and 70% （Kcp3））. Results indicated that lower irrigation quantities provided statistically lower yield and yield components. The highest seasonal fruit yield （80.0 t ha-1） was determined in the Kcp1 treatment, which applied the highest volume of irrigation water （452.9 mm）. The highest early fruit yield, average fruit weight and fruit diameter, length and number per plant were also determined in the Kcp1 treatment, with values of 7.25 t ha-1, 264.1 g, 5.49 cm, 19.95 cm and 10.92, respectively. Although the IWUE value was the highest in the Kcp1 treatment （176.6 kg ha-1 mm-1）, it was statistically similar to the value for Kcp3 treatment （157.1 kg ha-1 mm-1）. Total phenolic content and antioxidant activity of fruits was higher in the Kcp1 （44.27 μg gallic acid equivalents （GAE） mg-1 fresh sample） and in the Kcp2 （84.75%） treatments, respectively. Major （Na, N, P, K, Ca, Mg and S） and trace （Fe, Cu, Mn, Zn and B） mineral contents of squash fruits were the highest in the Kcp2 treatment, with the exception of P, Ca and Cu. Mineral contents and total phenolic content were signiifcantly affected by irrigation quantities, but antioxidant activity was not affected. It can be concluded that the Kcp1 treatment was the most suitable for achieving higher yield and IWUE. However, the Kcp2 treatment will be the most suitable due to the high fruit quality and relatively high yield in water shortage conditions.     

Short Response of Spring Wheat to Tillage,Residue Management and Split Nitrogen Application in a Rice-Wheat System

A ifeld experiment was conducted to study the impact of tillage, crop residue management and nitrogen （N） splitting on spring wheat （Triticum aestivum L.） yield over 2 yr （2010-2012） in a rice （Oryza sativa L.）-wheat system in northwestern Pakistan. The experiment was conducted as split plot arranged in randomized complete blocks design with three replications. Treatments comprised six tillage and residue managements：zero tillage straw retained （ZTsr）, zero tillage straw burnt （ZTsb）, reduced tillage straw incorporated （RTsi）, reduced tillage straw burnt （RTsb）, conventional tillage straw incorporated （CTsi）, and conventional tillage straw burnt （CTsb） as main plots and N （200 kg ha-1） was applied as split form viz., control （no nitrogen＆no splitting, N0S0）;2 splits of total N, half at sowing and half at the 1st irrigation （i.e., 20 d after sowing （DAS）） （NS1）;3 splits of total N, 1/3 at sowing, 1/3 at the 1st irrigation, and 1/3 at the 2nd irrigation （NS2）;4 splits of total N, 1/4 at sowing, 1/4 at the 1st irrigation, 1/4 at the 2nd irrigation （45 DAS）, and 1/4 at the 3rd irrigation （70 DAS） （NS3）;and 4 splits of total N, 1/4 at the 1st irrigation, 1/4 at the 2nd irrigation, 1/4 at the 3rd irrigation, and 1/4 at the 4th irrigation （95DAS） （NS4） as sub plots. The results showed that the most pikes m-2, grains/spike, 1 000-grain weight, grain yield, and N use efifciency （NUE） were obtained at zero tillage, straw retained and 4 splits application of total N （i.e., at sowing 20, 45 and 70 d after sowing）. The results indicated that ZTsr with application of 200 kg N ha-1 in 4 equal splits viz. at sowing 20, 45 and 70 d after sowing is an appropriate strategy that enhanced wheat yield （7 436-7 634 kg ha-1） and N efifciency （28.6-29.5 kg kg-1） in rice-wheat system.     

Complete Genome Sequence of Mycoplasma ovipneumoniae Strain NM2010, Which Was Isolated from a Sheep in China

Mycoplasma ovipneumoniae, a kind of mycoplasma bacteria, commonly infects the respiratory tract causing respiratory disease in sheep and goats worldwide. Here, the complete genome sequence of M. ovipneumoniae strain NM2010 isolated from a sheep in China was reported for the ifrst time.     

Effects of Dietary Fiber and Grit on Performance,Gastrointestinal Tract Development,Lipometabolism, and Grit Retention of Goslings

This study investigated effects of dietary ifbre and grit on growth performance, gastrointestinal tract （GIT） development, serum indexes and grit retention of Sichuan white goslings in China. The experiment was a 3×2 factorial design consisting of dietary crude ifbre （CF） at 2.0, 4.0 and 6.0%levels by adding grass meal and with or without grit supplementation. A total of 720 1-d-old male goslings were randomly allocated into 36 pens of 20 goslings/pen （n=6 pens/treatment）. At 21 d of age, GIT, blood samples and grit in gizzards were collected. Body weight （BW）, empty BW （without GIT）, average daily gain （ADG） and average daily feed intake （ADFI） were affected by CF levels and the medium （4.0%） had the highest value. Adding grit improved BW, empty BW, ADG and ADFI. CF affected relative weight or length of proventriculus, gizzard and duodenum. Increasing CF levels decreased jejunum villus height and affected caecal crypt depth and villus height to crypt depth ratio. Higher CF levels decreased serum triglyceride. Adding grit increased relative weight of abdominal fat. Grit addition increased grit weight in gizzard and the ratio of grit in 0.45-1 and 1-2 mm while decreasing the ratio of grit〈0.45 and 2-3 mm. In conclusion, moderate CF level and grit addition increased performance of goslings without interaction. Dietary CF levels affected the development of upper GIT and serum triglyceride. Grit needed by goslings should be less than 2 mm. Goslings’ gizzard would accumulate grit from feed when other grit could not be acquired.     

Fine Mapping and Candidate Gene Analysis of Resistance Gene RSC3Q to Soybean mosaic virus in Qihuang 1

Soybean mosaic virus （SMV） disease is one of the most destructive viral diseases in soybean （Glycine max （L.） Merr.）. SMV strain SC3 is the major prevalent strain in huang-huai and Yangtze valleys, China. The soybean cultivar Qihuang 1 is of a rich resistance spectrum and has a wide range of application in breeding programs in China. In this study, F1, F2 and F2：3 from Qihuang 1×nannong 1138-2 were used to study inheritance and linkage mapping of the SC3 resistance gene in Qihuang 1. The secondary F2 population and near isogenic lines （nILs） derived from residual heterozygous lines （RhLs） of Qihuang 1×nannong 1138-2 were separatively used in the ifne mapping and candidate gene analysis of the resistance gene. Results indicated that a single dominant gene （designated RSC3Q） controls resistance, which was located on chromosome 13. Two genomic-simple sequence repeat （SSR） markers BARCSOYSSR_13_1114 and BARCSOYSSR_13_1136 were found lfanking the two sides of the RSC3Q. The interval between the two markers was 651 kb. Quantitative real-time PCR analysis of the candidate genes showed that ifve genes （Glyma13g25730, 25750, 25950, 25970 and 26000） were likely involved in soybean SMV resistance. These results would have utility in cloning of RSC3Q resistance candidate gene and marker-assisted selection （MaS） in resistance breeding to SMV.     

Haplll of TaSAP1-A1, a Positively Selected Haplotype in Wheat Breeding

Stress-associated protein （SAP） has functions in maintaining plant cell elongation, embryo development and response to abiotic stresses. TaSAP1-A1, one of the Triticum aestivum SAP1 （TaSAP1） members located on wheat chromosome 7A was isolated for polymorphism analysis. HaplII of TaSAP1-A1 was found significantly associated with thousand-grain weight （TGW） in multiple environments. In this study, HaplII also made a positive contribution to TGW in Population 2. The distribution of TaSAP1-A1 HapIII was tracked among varieties released in different years and geographical environments of China. The frequency of HapIII showed an increasing trend during the breeding process in two different populations. The ItapIII was gradually selected and applied from 6.36% in landraces to 13.50% in modem varieties. These results exhibited that TaSAP1-A1 HapIII was positively selected during wheat breeding, which is beneficial for grain-yield improvement. The preferred HapIIl was initially selected and applied in the higher latitude areas of China in accord with the long day season and longer grain filling stage in these areas. Moreover, the frequency of HaplII in recent modem varieties was still quite low （19.29-26.67%）. It indicated a high application potential of TaSAP1-A 1 HapIII for improving grain yield in wheat breeding.     

Molecular Mapping of a Stripe Rust Resistance Gene YrH9020a Transferred from Psathyrostachys huashanica Keng on Wheat Chromosome 6D

Stripe rust （yellow rust）, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most devastating diseases of wheat throughout the world. H9020-1-6-8-3 is a translocation line originally developed from interspeciifc hybridization between wheat line 7182 and Psathyrostachys huashanica Keng and is resistant to most Pst races in China. To identify the resistance gene（s） in the translocation line, H9020-1-6-8-3 was crossed with susceptible cultivar Mingxian 169, and seedlings of the parents, F1, F2, F3, and BC1 generations were tested with prevalent Chinese Pst race CYR32 under controlled greenhouse conditions. The results indicated that there is a single dominant gene, temporarily designated as YrH9020a, conferring resistance to CYR32. The resistance gene was mapped by the F2 population from Mingxian 169/H9020-1-6-8-3. It was linked to six microsatellite markers, including Xbarc196, Xbarc202, Xbarc96, Xgpw4372, Xbarc21, and Xgdm141, lfanked by Xbarc96 and Xbarc202 with at 4.5 and 8.3 cM, respectively. Based on the chromosomal locations of these markers and the test of Chinese Spring （CS） nullitetrasomic and ditelosomic lines, the gene was assigned to chromosome 6D. According to the origin and the chromosomal location, YrH9020a might be a new resistance gene to stripe rust. The lfanking markers linked to YrH9020a could be useful for marker-assisted selection in breeding programs.     

Species Composition,Richness and Aboveground Biomass of Natural Grassland in Hilly-Gully Regions of the Loess Plateau,China

In order to study the characteristics of species composition, richness and aboveground biomass of natural grasslands, and then ifnd out the relations between species richness and aboveground productivity of the communities and possible mechanisms to form the relations, four typical grassland communities （Artemisia capillaries （AC）, Thymus quinquecostatus （TQ）, Stipa bungeana （SB） and Stipa grandis （SG）） along with a succession sequence in hilly-gully regions of the Loess Plateau, China, were investigated by ifeld survey and laboratory analysis. The results were summarized as follows：Different succession stages had different species compositions as well as different proportions of plant life forms and photosynthetic types, and Asteraceae, Poaceae and Leguminosae were their dominant species as well as their dominant perennial herb species;and different succession stages had signiifcantly different species richness and aboveground biomasses. There were many relation patterns （linear positive correlation, unrelated relations and unimodal relations） between the species richness and aboveground biomass in different succession stages and a signiifcant unimodal relation between the species richness and aboveground biomass in all the grassland communities and the highest species diversity appeared at a moderate level of productivity. The results suggest the unimodal relations in all the grassland communities are accumulative results of the relations in each succession stage.     

Inlfuence of Host Shift on Genetic Differentiation of the Oriental Fruit Fly,Bactrocera dorsalis

Invasion of the oriental fruit lfy, Bactrocera dorsalis, into new niches containing different food sources （a process referred to as host shift）, may cause population genetic differentiation and sympatric speciation. To attempt to infer that experimentally, test populations were established by transferring a subset of the original populations, which had been grown on banana for many generations, onto navel orange, and then subculturing the navel orange population and banana population for at least 20 generations. Four pairs of SSR primers with high polymorphism on laboratory strains were used to detect population genetic differentiation. All six tested populations （the 5th, 10th and 15th generations of B. dorsalis fed on banana and navel orange, respectively） were found to have low genetic diversity. Furthermore, the genetic diversity of the navel orange populations was found to decline after being crossed for several generations. Populations initially were deviated from Hardy-Weinberg equilibrium, however, equilibrium was achieved with increasing numbers of generations in both of the host populations. Limited gene lfows were found among the six populations. The Nei’s standard genetic distances between the two host populations of the same generation were initially low, but increased with generation number. Genetic distances between banana and navel orange populations of the same generation were lower than genetic distances between different generations grown on the same host plant. Analysis of molecular distance （AMOVA） results based on generation groups and host groups demonstrated that genetic variation among generations was greater than that between the two host populations. The results indicated that population genetic differentiation occurred after the host shift, albeit at low level. Biogeography and taxonomy of the B. dorsalis complex revealed that speciation of B. dorsalis might be tightly associated with host shift or host specialization of B. dorsalis following dispersal.