[The influences of different vegetation ecosystems on heavy metals in soil in semi-arid region].

The ecological environment of semi-arid regions in China is fragile, and the situation of protecting environment is grim. So it is urgent to strengthen environment protection and ecological construction in semi-arid region. Four different vegetation ecosystems were selected in semi-arid region with Wuchuan County in Inner Mongolia as a case study: the bushes, the trees, the alfalfa land, and artificially mixed sowing grassland. The soil was sampled and carried to laboratory for analysis of the content of lead, cadmium and chromium in the soil in May and September (the start of the growing season and the end of the growing season). It was showed that among the four different ecosystems, the lead and cadmium contents in the soil were significantly different in variability, while the chromium is not significant. And the changing rate trend of the content of lead and cadmium is consistent: the contents of the both elements in May are higher than in September obviously. The degree of the order of the content change is also very close. The biggest of the change scope is from the bushes and artificially mixed sowing grassland, followed by the alfalfa land, and finally the trees. Therefore, it is more favorable to plant shrubs and grass for absorbing heavy metals in the soil in the common local vegetation eco-system.

[Attempting to enhance the efficiency of T-DNA insertional mutant application in rice].

T-DNA tagging method is a high throughput system for identifying and cloning novel genes from T-DNA-inserted mutant population created via genetic transformation by Agrobacterium tumefaciens. However, the efficiency of using T-DNA-inserted mutant population to clone genes in rice was much lower than in Arabidopsis. In this study, a rice tagged line with two copies of T-DNA segments inserted independently to each other was screened out via a series of verification tests, including the co-segregated analysis between the mutated character and the sequence of T-DNA or the genomic sequence flanking inserted T-DNA. From this tagged line, two inserted incidents were separated from the progeny population of a plant heterozygous in two tagged sites, and some plants with the target trait and one of the inserted incidents were obtained, which were important basic materials for the subsequently co-segregated analysis between the mutated character and the sequence of inserted T-DNA, and for cloning the mutant gene in future. Based on this study, we have some thoughts about the gene cloning from the T-DNA tagged lines with more than one inserted sequence independently and put forward to discuss with colleagues.

Screening for and genetic analysis on T-DNA-inserted mutant pool in rice.

T-DNA tagging technique has provided a powerful strategy for identifying new functional genes in plants, and the key for success is the discovery of T-DNA-inserted mutants with changed phenotype. In this study, we screened 4,416 rice T1 tagged lines generated by enhancer trap system integrated with GLL4/VP16-UAS elements from two transformed parents, ZH11 and ZH15. We found many lines showed obvious morphological mutations, including two types--fake-homozygous mutation and separating mutation. The mutation phenotype was related to 14 kinds of trait such as plant height, heading date, leaf shape, leaf color, tiller number, panicle shape, spikelet number, grain shape, disease-like mutation, male sterility, awn, and so on. Among them, plant height, heading date, leaf color and male sterility had a comparatively high mutation frequency (over 1%). The mutation frequency of plant height and leaf color had no significant change between different years or transformed parents, but the frequency of heading date and male sterility varied greatly, suggesting that environment had a great effect on the expression of latter two traits. By conducting continuously co-segregating analyses in T1 and T2 generation, we identified 3 T-DNA-inserted mutants with malformed panicle or spikelets, which would provide a base for cloning correlative functional genes. At the same time, we selected randomly 42 lines with mutation phenotype and obtained 40 flanking sequences from 39 tagged lines by plasmid rescue or TAIL-PCR, of which, 26 were vector backbone sequence, 14 had good identity to rice genome sequence. The BlastN result showed the T-DNA preferentially integrated into protein-coding region in plants.

[One major QTL mapping and physical map construction for rolled leaf in rice].

A clonally propagated F2 population (F2CP), derived from the rice cross of Qimiaoxiang/91SP068, was used to map rolled leaf QTLs. As the parent Qimiaoxiang is an unrolling leaf variety, while 91SP068 is a medium rolling variety with about 34% rolling index. One major QTL, rl8, which came from 91SP068, was mapped between two flanking SSR markers, RM6954 and RM6841, on chromosome 5, with genetic distance 3.8 cM, and was 1.0 cM away from RM6954. Its additive effect estimated by composite interval mapping (CIM) was 9.61 in 2002 and 6.23 in 2003, and the dominance effect was also different between two years, -1.19 in 2002 and -4.44 in 2003, respectively. It explained about 20% - 33% of the total phenotypic variation between two years. Furthermore, an integrated physical and genetic map encompassing rl8 region was constructed, and the physical distance of the interval was 542 kb, and the ratio of physical to genetic distance was 144 kb/cM. Based on this research, fine mapping of rolled leaf QTLs will not only facilitate the map-based cloning of the gene itself, but also improve the efficiency of marker-assisted selection in plant breeding.

[Identification and marker-assisted selection of two major quantitative genes controlling rice sheath blight resistance in backcross generations].

We constructed an F2 clonal population of intercross,Teqing/Lemont, and identified two quantitative trait loci (QTLs) contributing to rice sheath blight resistance on chromosome 9 and 11. The two QTLs were qSB-9 and qSB-11, respectively. From the population, three clonal lines were selected by markers' band types on both sides of these two QTLs, qSB-9 and qSB-11. Two were double-susceptible parent with homozygous susceptible alleles of these two loci,and the other was named as double-resistant parent,of which these two loci were all homozygous resistant alleles. These parents were separately backcrossed to recurrent parents, Teqing or Lemont. From BC2F1, marker-assisted selection was conducted in each proceeding generation and all back-crossed plants in BC2F1 and BC4F1 were inoculated by short toothpicks incubated with a strain, RH-9 of the fungus for identification of the resistance. Results suggested that these two QTLs were selected effectively in each backcross generation and their positions were also verified in identification of resistance to rice sheath blight. In seedling nursery of BC3F2 population, plants were selected through marker-assisted selection, and were separately mixed as homozygous lines of double-susceptible alleles on the background of Teqing, double-susceptible and double-resistant on the background of Lemont. The homozygous lines and their recurrent parents were simultaneously planted on experiment fields of Agriculture Collage of Yangzhou University and Lixiahe District Institute of Agricultural Science. The inoculation was performed by a random-block test with two replicates at each site. The results indicated that 1) The difference of sheath blight disease development was highly significant among materials under the same genetic background,and the order of disease seriousness among different homozygous lines were: double-susceptible line on the background of Lemont > double-susceptible line on the background of Teqing > Lemont > Teqing > double-resistant line on the background of Lemont; 2) When the resistant allele of qSB-9 or qSB-11 solely existed in a plant, its disease rating was reduced about 1.2 score, and 2.0 score when they simultaneously existed on the background of Lemont; 3) No significant interaction between the two QTLs controlling sheath blight resistance and environments was found. These studies have laid a strong groundwork in operation and application, of these QTLs contributing to rice sheath blight resistance.

[Mapping QTLs for horizontal resistance to sheath blight in an elite rice restorer line, Minghui 63].

This study was conducted with a recombinant inbred line (RILs) population consisting of 240 recombination lines, derived from an elite combination, Zhenshan 97B x Minghui 63. The RILs and their parents were grown in a randomized complete design with two replications in the years of 1999 and 2000. Sheath blight response ratings for the population and their parents were identified by an improved method of inoculation, which was carried out with short woody toothpicks incubated with a Rhizoctonia solani strain, RH-9, and inserted the third sheath in the late tillering/green ring stage of growth. A linkage map was constructed from the RILs. The QTL mapping of sheath blight resistance was carried out by the method of interval QTL mapping. Two QTLs for sheath blight resistance were detected in each year, and were located on chromosome 5 and chromosome 9, respectively. The QTL for sheath blight resistance on chromosome 5 was flanked by markers C624 and C246 on the basis of 1999 data, and by markers C246 and RM26 using 2000 data. The 1-LOD-confidence intervals of QTLs for sheath blight resistance on chromosome 5 detected in two years greatly overlapped with each other, and the peak of the 1-LOD-confidence intervals were approximately the same site. This suggested that the QTL for resistance on chromosome 5 detected in 1999 was probably the same as the QTL detected in 2000. The QTL for sheath blight resistance on chromosome 9 was located on the marker interval of C472-R2638 in term of 1999 data, and on the interval of RM257-RM242 based on 2000 data, and the two intervals were 9.7 cM away from each other. Based on the effect analysis of QTLs for resistance, the genotype of MH63 had negative additive effects or reduced sheath blight rating.