Fine mapping of qSB-11(LE), the QTL that confers partial resistance to rice sheath blight.

Sheath blight (SB), caused by Rhizoctonia solani kühn, is one of the most serious global rice diseases. No major resistance genes to SB have been identified so far. All discovered loci are quantitative resistance to rice SB. The qSB-11(LE) resistance quantitative trait locus (QTL) has been previously reported on chromosome 11 of Lemont (LE). In this study, we report the precise location of qSB-11 (LE) . We developed a near isogenic line, NIL-qSB11(TQ), by marker-assisted selection that contains susceptible allele(s) from Teqing (TQ) at the qSB-11 locus in the LE genetic background. NIL-qSB11(TQ) shows higher susceptibility to SB than LE in both field and greenhouse tests, suggesting that this region of LE contains a QTL contributing to SB resistance. In order to eliminate the genetic background effects and increase the accuracy of phenotypic evaluation, a total of 112 chromosome segment substitution lines (CSSLs) with the substituted segment specific to the qSB-11 (LE) region were produced as the fine mapping population. The genetic backgrounds and morphological characteristics of these CSSLs are similar to those of the recurrent parent LE. The donor TQ chromosomal segments in these CSSL lines contiguously overlap to bridge the qSB-11 (LE) region. Through artificial inoculation, all CSSLs were evaluated for resistance to SB in the field in 2005. For the recombinant lines, their phenotypes were evaluated in the field for another 3 years and during the final year were also evaluated in a controlled greenhouse environment, showing a consistent phenotype in SB resistance across years and conditions. After comparing the genotypic profile of each CSSL with its phenotype, we are able to localize qSB-11 (LE) to the region defined by two cleaved-amplified polymorphic sequence markers, Z22-27C and Z23-33C covering 78.871 kb, based on the rice reference genome. Eleven putative genes were annotated within this region and three of them were considered the most likely candidates. The results of this study will greatly facilitate the cloning of the genes responsible for qSB-11 (LE) and marker-assisted breeding to incorporate qSB-11 (LE) into other rice cultivars.

An inactivation kinetics model for Campylobacter jejuni on chicken meat under low-temperature storage.

Campylobacter jejuni is susceptible to low temperatures. Freezing and chilling are effective interventions for reducing the occurrence of C. jejuni on poultry meat. The survival rates of three C. jejuni strains (ATCC33560, JR0706-2, and ALM-80) inoculated onto chicken meat samples were measured at -20°C and 4°C, and the survival curves of these three strains were determined. The results showed that the number of surviving cells decreased by 3.16, 2.87, and 3.14 log colony-forming unit (CFU)/g, respectively, at -20°C during the 55-day storage period. The survival curves showed that the mean inactivation speeds were slow in the initial 20 days of storage at -20°C, dropped rapidly between 25 and 45 days, and reached a plateau in reduction between 45 and 55 days. The number of surviving cells during the 10-day storage period at 4°C decreased by 3.47, 3.35, and 3.51 log CFU/g, respectively. The mean inactivation speeds at 4°C were 0.347, 0.355, and 0.439 log CFU/day. There were some differences in the inactivation speeds of the three strains of C. jejuni, but there was no significant difference in the trend of inactivation among the strains (p>0.05). Only the seven parameters are different for the strains originating from diverse sources when the inactivation trend was determined using a formula. Data-fitting software, MATLAB, was used to fit the survival rates data. The results showed that the established inactivation kinetics function fit the data well for the three different strains stored at -20°C and 4°C. This study revealed the inactivation kinetics for three C. jejuni strains on chicken meat during low-temperature storage and provided useful information for C. jejuni risk management.

Quantitative trait loci for resistance to fusarium head blight in a Chinese wheat landrace Haiyanzhong.

Fusarium head blight (FHB) of wheat causes not only significant reduction in grain yield and end-use quality, but also the contamination of the grain with mycotoxins that are detrimental to human and animal health after consumption of infected grain. Growing resistant varieties is an effective approach to minimize the FHB damage. The Chinese wheat landrace Haiyanzhong (HYZ) shows a high level of resistance to FHB. To identify quantitative trait loci (QTL) that contribute to FHB resistance in HYZ, 136 recombinant inbred lines (RIL) were developed from a cross of HYZ and Wheaton, a hard spring wheat cultivar from the USA. The RIL and their parents were evaluated for percentage of scabbed spikelets (PSS) in both greenhouse and field environments. Five QTL were detected for FHB resistance in HYZ with one major QTL on 7DL. The 7DL QTL peaked at SSR marker Xwmc121, which is flanked by the SSR markers Xcfd46 and Xwmc702. This QTL explained 20.4-22.6% of the phenotypic variance in individual greenhouse experiments and 15.9% in a field experiment. Four other minor QTL on 6BS (two QTL), 5AS and 1AS each explained less than 10% of the phenotypic variance in individual experiments. HYZ carried the favorable alleles associated with FHB resistance at the QTL on 7DL, 6BS and 5AS, and the unfavorable allele at the QTL on 1AS. The major QTL on 7D can be used to improve the FHB resistance in wheat breeding programs and add diversity to the FHB resistance gene pool.

Genetic analysis of starch paste viscosity parameters in glutinous rice (Oryza sativa L.).

Starch paste viscosity plays an important role in estimating the cooking, eating, and processing quality of rice. The inheritance of starch paste viscosity in glutinous rice remains undefined. In the present study, 118 glutinous rice accessions were collected, and the genotypes of 17 starch synthesis-related genes (SSRG) were analyzed by using 43 gene-specific molecular markers. Association analysis indicated that 10 of 17 SSRGs were involved in controlling the rapid visco analyzer (RVA) profile parameters. Among these, the PUL gene was identified to play an important role in control of peak viscosity (PKV), hot paste viscosity (HPV), cool paste viscosity (CPV), breakdown viscosity (BDV), peak time (PeT), and paste temperature (PaT) in glutinous rice. Other SSRGs involved only a few RVA profile parameters. Furthermore, interactions between SSRGs were found being responsible for PeT, PaT, and BDV. Some of the RVA parameters, including PKV, HPV, CPV, CSV, and PaT, were mainly governed by single SSRG, whereas other parameters, such as BDV, SBV, and PeT, were controlled by a few SSRGs, functioning cooperatively. Further, three near-isogenic lines (NIL) of a japonica glutinous cv. Suyunuo as genetic background, with PUL, SSIII-1, and SSIII-2 alleles replaced with those of indica cv. Guichao 2, were employed to verify the genetic effects of the various genes, and the results were consistent with those obtained from the association analysis. These findings indicated that starch paste viscosity in glutinous rice had a complex genetic system, and the PUL gene played an important role in determining the RVA profile parameters in glutinous rice. These results provide important information for potentially improving the quality of glutinous rice.

Molecular evolution and functional divergence of HAK potassium transporter gene family in rice (Oryza sativa L.).

The high-affinity K(+) (HAK) transporter gene family is the largest family in plant that functions as potassium transporter and is important for various aspects of plant life. In the present study, we identified 27 members of this family in rice genome. The phylogenetic tree divided the land plant HAK transporter proteins into 6 distinct groups. Although the main characteristic of this family was established before the origin of seed plants, they also showed some differences between the members of non-seed and seed plants. The HAK genes in rice were found to have expanded in lineage-specific manner after the split of monocots and dicots, and both segmental duplication events and tandem duplication events contributed to the expansion of this family. Functional divergence analysis for this family provided statistical evidence for shifted evolutionary rate after gene duplication. Further analysis indicated that both point mutant with positive selection and gene conversion events contributed to the evolution of this family in rice.

Molecular evolution of the CPP-like gene family in plants: insights from comparative genomics of Arabidopsis and rice.

CPP-like genes are members of a small family which features the existence of two similar Cys-rich domains termed CXC domains in their protein products and are distributed widely in plants and animals but do not exist in yeast. The members of this family in plants play an important role in development of reproductive tissue and control of cell division. To gain insights into how CPP-like genes evolved in plants, we conducted a comparative phylogenetic and molecular evolutionary analysis of the CPP-like gene family in Arabidopsis and rice. The results of phylogeny revealed that both gene loss and species-specific expansion contributed to the evolution of this family in Arabidopsis and rice. Both intron gain and intron loss were observed through intron/exon structure analysis for duplicated genes. Our results also suggested that positive selection was a major force during the evolution of CPP-like genes in plants, and most amino acid residues under positive selection were disproportionately located in the region outside the CXC domains. Further analysis revealed that two CXC domains and sequences connecting them might have coevolved during the long evolutionary period.

Genomewide comparative phylogenetic and molecular evolutionary analysis of tubby-like protein family in Arabidopsis, rice, and poplar.

Tubby-like proteins, which are characterized by a highly conserved tubby domain, play an important role in the maintenance and function of neuronal cells during postdifferentiation and development in mammals. In additional to the tubby domain, most tubby-like proteins in plants also possess an F-box domain. Plants also appear to harbor a large number of TLP genes. To gain insight into how TLP genes evolved in plants, we conducted a comparative phylogenetic and molecular evolutionary analysis of the tubby-like protein gene family in Arabidopsis, rice, and poplar. Genomewide screening identified 11 TLP genes in Arabidopsis, 14 in rice, and 11 in poplar. Phylogenetic trees, domain organizations, and intron/exon structures classified this family into three subfamilies and indicated that species-specific expansion contributed to the evolution of this family in plants. We determined that in rice and poplar, the tubby-like protein family had expanded mainly through segmental duplication events. Tissue-specific expression analysis indicated that functional diversification of the duplicated TLP genes was a major feature of long-term evolution. Our results also demonstrated that the tubby and F-box domains had co-evolved during the evolution of proteins containing both domains.

Comparative study of SBP-box gene family in Arabidopsis and rice.

SBP-box proteins are plant-specific putative transcription factors, which contain highly conserved SBP domain and could bind specifically to promoters of the floral meristem identity gene SQUAMOSA and its orthologous genes to regulate their expressions. In this study, 17 non-redundant SBP-box genes in Arabidopsis genome and 19 in rice genome were identified by using the known SBP domain sequences as queries. The phylogenetic analysis suggested that the main characteristics of this family might have been in existence before the split of Arabidopsis and rice, and most SBP-box genes expanded in a species-specific manner after the split of monocotyledon and dicotyledon. All the SBP-box proteins were classified into 9 subgroups based on the phylogenetic tree, where each group shared similar motifs and the orders of the motifs in the same group were found almost identical. Analysis of nonsynonymous and synonymous substitution rates revealed that the SBP domain had gone through purifying selection, whereas some regions outside SBP domain had gone through positive or relaxed purifying selection. The expression patterns of the SBP-box genes were further investigated by searching against the EST database. Results showed that the Arabidopsis SBP-box genes are expressed chiefly in flowers, leaves, roots and seeds, while those in rice mainly in flowers and callus.

Divergence of reproductive phenology under climate warming.

Because the flowering and fruiting phenology of plants is sensitive to environmental cues such as temperature and moisture, climate change is likely to alter community-level patterns of reproductive phenology. Here we report a previously unreported phenomenon: experimental warming advanced flowering and fruiting phenology for species that began to flower before the peak of summer heat but delayed reproduction in species that started flowering after the peak temperature in a tallgrass prairie in North America. The warming-induced divergence of flowering and fruiting toward the two ends of the growing season resulted in a gap in the staggered progression of flowering and fruiting in the community during the middle of the season. A double precipitation treatment did not significantly affect flowering and fruiting phenology. Variation among species in the direction and magnitude of their response to warming caused compression and expansion of the reproductive periods of different species, changed the amount of overlap between the reproductive phases, and created possibilities for an altered selective environment to reshape communities in a future warmed world.

Molecular marker analysis and genetic basis for sterility of typical indica/japonica hybrids.

To explore the genes differentiated between typical indica and japonica varieties, two typical indica/japonica varieties, Balilla (japonica) and Nantehao (NTH, indica), were selected to construct genetic populations based on the widely surveying for spikelet and pollen fertility of 90 indica/japonica F1 hybrids, which also were used as the wide compatability testers. In order to analyze the genes (QTLs) related to spikelet and pollen fertility, two reciprocal backcross populations Balilla/NTH//Balilla and Balilla/NTH//NTH were constructed and the spikelet and pollen fertility of each individuals were assessed. In both populations, two traits all appeared distorted normal distribution, but in the first population, they forwarded to low-level fertility type, the later population, forwarded to high-level fertility type relatively. The results indicated that both of male and female gametophytes of Balilla/NTH hybrids were partial sterile. Then we analyzed the SSR marker genotype of each individuals of Balilla/NTH//Balilla population containing 142 individuals, and constructed a SSR linkage map, in which, there were 108 information markers distributing on all 12 chromosomes equably, average marker distance was about 11.9 cM. Therefore the linkage map was qualified for QTL analysis. Two methods were employed to conduct QTLs analysis, i.e., single marker analysis and interval mapping. According to single marker analysis, 17 and 12 markers were found significantly responsible for spikelet and pollen fertility, respectively. And further study by means of MAPMAKER/QTL software, for spikelet fertility trait, two QTLs were detected, qSPTF1 on chromosome 1 and qSPTF6 on chromosome 6, and their additive effect were 13.501 and -16.414, respectively. According to previous studies, qSPTF6 was deduced to be the same locus as S-5. For pollen fertility, qPLLN7 on chromosome 7 and qPLLN9 on chromosome 9 were detected, and their additive effects were -12.003 and -11.012, respectively. Because the QTLs detected cannot explain completely the total variance of mapping population, other genetic factors must be existed to be responsible for spikelet and pollen partial sterility. Hence we let two random markers as putative covariates, and divide the 142 individuals into four groups according to the two marker genotypes, then the average values of spikelet and pollen fertility of each groups were calculated for two-way ANOVA (analysis of variance). The results indicated that there existed strong interaction for both spikelet fertility and pollen fertility. At a significance level of 0.005, there over 61 and 51 pairs loci interactions detected playing an important role in spikelet and pollen sterility expression, respectively. These results indicated that epistasis also was one of major genetic components controlling indica/japonica hybrid sterility.

[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.