Using a soybean transgenic line which is resistant to glyphosate and its receptor material as positive control and negative control respectively,five soybean cultivars partially survived after the treatment with spraying of glyphosate were examined by both the test strip and PCR methods.The results showed that the CP4 EPSPS gene was detected in these soybean materials by both methods,at either protein or DNA level.And the test strip detection was consistent with PCR detection.Further application of these two techniques was discussed.
The combining ability of seven yield traits were analysed with the F_(4∶5) lines of 16 combinations by 4×4 incomplete diallel crossing.The variation of all traits among female parents and four traits,pods and seeds weight per plant,100-pod and-seed weight among male parents were significant at 1% probability level,and shelling percentage of male parents was significant at 5% probability level.The general combining ability(GCA) of 963-4-1 and the special combining ability(SCA) variance of 904-79 among the maternal lines were the highest on pod and seed weight per plant,pods and mature pods per plant and shelling percentage.And the GCA of Zhonghua No.8 was the highest on pods and seeds per plant among the male parents and it's other traits performed well.The combination of 904-79 × Zhonghua(No.8) had the bigest SCA.The majority of high yielding lines which significantly over-yielded Tianfu No.9 was derived from 963-4-1× or 904-79× Zhonghua(No.8.) It was noticed that GCA or SCA of the lines in F_(4∶5) lines were inconsistent with these detected at earlier generation,therefore the overall performance of parental lines in several generations should be considered in peanut variety improvement.
HPLC was applied to analyze the profiles and contents of glucosinolates which were extracted from Brassica leaves with 70% methanol solvent by ultrasonic methods after freeze-drying.Results showed that the recoveries ranged from 94% to 103%,the range was 2.29μmol/g,which accorded with ISO9167 standard.The method was precise,simple and practical for leave glucosinolate determination.It could be widely used in quality and diseases resistance breeding for analysis of glucosinolates contents and profiles in Brassica leaves.
To establish high-yield direct-seeding cultivating technique of a rape cultivar Zhongshuang 9, a quadratic rotational regression design was used to study the effects of cultivating techniques on biological performance before winter and seed yield. A suitable cultivating technique for high yield (more than 3 000kg/hm~2) rapeseed production was developed. The correlation between plant biological indexes before winter and seed yield was discussed.
Two kinds of PGPR microbial agent were applied in rapeseed field and agronomic characters, economic traits, theoretical yield and sclerotinicse withered stalk ratio in different growth stage were examined to study the growth promoting function and biological control effect. The result showed that PGPR microbial agent has obvious growth-promoting effect on rapeseed and could significantly improve its agronomic characters, especially the number of siliquae per plant and yield. PGPR microbial agent also has obvious biocontrol effect and decreased the infection ratio of stem rot caused by S.sclerotiorum.There existed different effects between different types of PGPR microbial agent, the effect of seed-coating type and YUMI compound liquid type was the best to promote growth and biological control for rape, and YUMI liquid type was the second.
Transgenic peanut lines TP-1 and TP-7 harboring coat protein (CP) gene of peanut stripe virus ( PStV) were used to study the effect of transgene on Rhizobium. Results showed that there was no significant difference on average Rhizobium nodule numbers and weight between transgenic and non-transgenic peanut plants at various growing stages. One hundred Rhizobium samples were isolated from nodules of transgenic and non-transgenic peanut plants, respectively. PCR test revealed that there was no CP gene in Rhizobium of transgenic peanut plants. The results indicated that the possibility of CP gene dispersed from transgenic peanut to Rhizobium was very low.
Three hundred and twenty bacteria isolates were isolated from rhizospere and phyllophere of rapeseed, 18 of the isolates were found suppressing Scleotinia sclerotiorum mecylium growth on the PDA plate, among them Y1 was found efficiently suppress Scleotinia sclerotiorum. Y1 was also found efficiently suppress Scleotinia sclerotiorum in biocontrol test on rapeseed detached leaves, seedling in the greenhouse and field plot. The efficacy of 92% was obtained in biocontrol test in the greenhouse and field plot. This bacteria isolate was identified as Bacillus subtilis.
Analytical methods for aflatoxin B_1 were briefly reviewed in this paper. Recent development of new fast test techniques for aflatoxin B_1 analysis in grains and oilseeds products based on immunoaffinity were introduced and discussed.
The Carboxyl-modified polyacrylamide microsphere was applied as the carrier. After the carboxyl group was activated by EDC, carboxyl-modified polyacrylamide microsphere combined the antibody of Aflatoxin B_1 by the covalence bond. The highest coupling efficiency reached 60.6% when the optimized buffer pH was between 6.0 and 6.5 with the reaction time of 17~20 hours and EDC amount of 5-15mg/10mg microsphere applied.
Phosphoenolpyruvate carboxylase(PEPase) plays an important role in the control of the ratio of the protein/lipid content in rape seed (Brassica napus). The cDNA fragment of PEPase gene was cloned from B.napus zhongshuang No.4 by RT-PCR. Sequence analysis of the fragment suggested that it possessed 98% homology with the PEPase gene(D14985). In order to down-regulate the expression of PEPase gene, two 400bp fragments were cloned into the Sap I site of RNAi vector pHBM1301 and two recombinant plasmids pHBM1301-PEP1 and pHBM1301-PEP2 were obtained.
Three genic male sterile lines and twenty seven core collections in sesame were used for measuring eleven quantitative traits, and the principal components, genetic distance, clustering and heterosis were analyzed. The results showed that the 30 genotypes could be classified into five clusters and the grouping of genotypes into clusters was not related to their geographic origin. The male sterile lines Ms01 and Ms03 were delimited into cluster Ⅰ and Ms02 into cluster Ⅲ. There was no clear relation between genetic distance and heterosis. The largest heterosis hybrid was Ms02×ZZM2918 with the seed yield increase of 22.97% over the control, Zhongzhi 10.
A soybean choloroplast DNA fragment named soy was isolated by PCR based on the published sequence (GeneBank X076075) and inserted into a fragment containing Amp~r and ColE1 ori from pBluescript SK(+) to generate soybean chloroplast expression vector pCS with an expression cassette containing three tandem genes (aadA, man and gfp). The function of the expression cassette was identified by plate qualitative assay and Western blotting in E.coli. The results showed that the multicistrons were expressed effectively through soybean chloroplast expression vector pCS.
Genetic distances between 3 double low self-incompatible lines and 22 pure line varieties from different geographic origins and performance of 66 hybrids produced by 3 SI lines and these varieties within a NCII mating design for yield traits were analyzed in this study. Results indicated that hybrids made from 3 SI lines and varieties from foreign countries showed the highest mid-parent heterosis for yield/plant, though the genetic differences between the SI lines and the male parents were not distinct. Generally, F_1 hybrid's days from emergence to maturity and from initial flowering to maturity were both between those of the two parents, but the former was closer to that of the serotinous parent, and the later was closer to that of the precocious one. Primary branches and their siliques contributed most not only to seed yield/plant, but also to heterosis of yield/plant. Therefore, the preferred candidate traits in genetic improvement, especially in application of heterosis in rapeseed, were primary branches and their siliques.
The weed control effects of paraquat and subsequent dynamics in untilled rape fields,as well as the influence of paraquat on the sunlight and crop yield were studied to evaluate the application prospect of paraquat and establish a weed management system in untilled rapeseed fields.The results showed that paraquat provided good control effects on weeds in untilled rapeseed fields,the overall effects ranged from 98.50% to 99.49% five days after treatment(DAT),decreased and maintained above 85% for 160 DAT with graduall decrease.The efficacy was higher on Alopecurus aequalis,Malachium aquaticum and Beckmannia syzigachne,than on Mazus japonicus.Paraquat application significantly improved the light conditions in rapeseed field and the light penetration rate increased with the dosage of herbicide.The application of paraquat had no effect on the plant height and 1000-seed weight,whereas the height of first branch,numbers of branches and siliquae were markedly influenced,resulting in a significant yield increase of rapeseed.
Nine hundred and eighty-one diseased specimens of soybean leaves were collected from Middle and Northern Huang Huai Region in 2002 and 2003,and 95 SMV isolates were obtained using biological purification and serology(DAS-ELISA).Based on symptomatic response of 27 differential hosts to these strains,Nannong 1138-2,Youbian 30,8101,Tiefeng 25,Davis,Bullfalo,Zaoshou 18,Kwanggyo,Qihuang 1 and Kefeng 1 were chosen as the differential hosts for the identification of the SMV strain groups in Northern Huang Huai Region of China.The 95 SMV isolates were classified into 10 strain groups.Five new strain groups were discovered and named SC-11~SC-15.In Northern Huang Huai Region,SC11 was preponderant,accounting for 21.1%.The second main strain group was the SC-8,accounting for 17.9%.The strain groups with the widest distribution were SC-11 and SC-13,which can be detected in all provinces of the region.
The restorer-maintainer relationship determination were performed by crossing the NCa,nap,pol,ogura cytoplasmic male sterile(CMS) lines with 104 accessions of different originals.Eighty out of the 104 accessions could restore the fertility of NCa CMS,and 80% of these restoring materials distributed mainly in China and Europe,whereas only 8 accessions could maintain the sterility of NCa CMS.For the pol,nap and ogura CMS,83 out of 104 accessions could maintain the sterility of pol CMS,while 99 accessions could restore the fertility of nap CMS,and 1 accession could restore the fertility of ogura CMS. The proportion of accessions which showed different fertility between NCa CMS and nap,pol,ogura CMS were 25%、89%、92%,respectively.NCa CMS may be classified into a CMS type different from nap,pol,ogura CMS by restorer-maintainer relationship determination.RFLP analysis of mitochondrial DNA from ogura、nap、 pol、NCa CMS with the enzyme/probe combinations of atp1/EcoR Ⅰ、atp1/Hind Ⅲ、atp1/BamHⅠ 、atp1/PstⅠ was conducted,and discovered significant diversity among the four CMS lines.It was proved that NCa CMS is definitely different from the other three CMS lines by mitochondrial polymorphism analysis.
Effects of salt stress with 100~300mmol/L NaCl concentrations on the emergence, growth and ionic distribution of oil sunflower seedlings were studied. Results showed that oil sunflower was highly tolerant to salt during seedling emergence. Different concentrations of NaCl (100~300mmol/L) didn’t decrease seedling emergence, but delayed seedling emergence. 100~300mmol/L NaCl inhibited remarkablely seedling growth and decreased dry matter accumulation rate. Na~+ was mainly accumulated in the stem and root of the seedlings, with little in the leaves. Cl~- was mainly distributed in stem and petiole, with lower content in the leaves and root. K~+ content in the leaves and petiole increased remarkably after exposion to salinity. Both absorption of S_(K,Na) in root and transportation of S_(K,Na) in the leaves and petiole were promoted remarkably. This suggested the high selectivity of K~+ in oil sunflower during the course of ion absorption and transportion. These were main reasons for the high salt-tolerance of oil sunflower.
In order to evaluate the food safety of anti-PEP transgenic rape Chaoyou No.1,acute toxicity and mutagenesis experiments were conducted.Results showed that the oil made from Chaoyou No.1 ranked as nontoxic class and no mutagenesis action was found.In a 90-day feeding test with rats,no marked differences in appearance,body weight,food intake,feed utilization rate,organ relative weight and hemogram of rates were observed between all treated groups and the control,and no pathological changes found in organs such as liver and kindney,with none effect dosage being 10.0ml/kg.
In 1998,a sterile mutant plant was observed in apetalous lines in B.napus L,and an apetalous mutant sterile(AMS) line was established from this plant.Studies showed that the sterility of this male sterile line was very complete,with male organs converting to female organs,and other morphologic characteristics were similar to those of apetalous fertile plants.779 test crosses were made with 549 materials including different varieties and maintainers,restorers and F_1 hybrids of different CMS systems.All restored AMS fertility completely.So the restoring resources were very wide.In 2002 and 2003,yield of 29 combinations from 100 combinations out-yielded CK(hybrid rapeseed variety Qinyou No.7) by 5.7%~23.5%.The level of yield increase was significant.When the stertile plants crossed with fertile plants in fertility segregation populations,1∶1 genetic segregation model appeared in the progeny.According to fertility segragation from other test cross and selfcrosses,the sterility was probably controlled by 4 pairs of recessive genes.The discovery of AMS not only provided special germplasm resource for studies on floral organ development mechanism,but also open up a new way in heteriosis utilization of rapeseed.
The antifungal ability of T.aureoviride against several strains of Fusarium spp.,which are pathogens of soybean root rot was studied in vitro by dual culture and treating with fermented culture filter.The effect of this strain on seedling root rot of soybean was also tested in pot experiment.Under microscope and TEM,the mycelia of the two fungi occurred across, entangle or parallel,and the mycelium of F.spp was destroyed by T.aureoviride.The fermented culture filter prolonged spore germination.Sorbitol,glucose and sucrose as carbon sources was better than other carbon sources in restraining the growth of pathogen mycelium.After application of T.aureoviride,the control effect of root rot reached 73.2%,with increased the dry matter weight and the height of soybean seedling.
