Yuxiang Zeng

Advances in Mapping Loci Conferring Resistance to Rice Sheath Blight and Mining Rhizoctonia solani Resistant Resources

Sheath blight (SB) caused by Rhizoctonia solani is one of the three major diseases of rice, and now has become the most severe disease causing rice yield loss in China. Breeding and use of varieties resistant to SB is crucial in controlling the disease, but the advances achieved have been limited due to the lack of highly SB-resistant rice germplasm. Genetic analysis revealed that the SB resistance in rice was a typical quantitative trait controlled by multi-genes. Although many QTLs conferring resistance to SB have been identified in recent years, most of the QTLs only showed small effects and few of them have been evaluated for utilization potential. Many R. solani-resistant resources have been found in wild rice species, microorganisms and other plant species. It is already known that the SB-resistance could be improved in transgenic rice plants by genetic transformation. This paper reviewed the genetic mapping of loci associated with resistance to rice SB, the evaluation of the potential of resistance QTLs, and the resistant resources found in various organisms besides rice. To develop SB-resistant rice varieties, it is important to develop and explore new resistant rice germplasms, fine map and evaluate resistance QTLs, and also to pay attention to various bio-resources showing resistance to R. solani.

Identification of QTLs for Rice Cold Tolerance at Plumule and 3-Leaf-Seedling Stages by Using QTLNetwork Software

Abstract A doubled haploid (DH) population consisted of 120 lines, derived from a cross between an indica variety, TN1, and a japonica variety, Chunjiang 06, was used to identify QTLs controlling rice cold tolerance at the plumule and 3-leaf-seedling stages by using the QTLNetwork software. The percentages of normal plumules after 4°C treatments for 7 d, 9 d, 11 d, and 14 d, as well as the cold stress tolerance index (CSTI) and the withering index (WI) of rice seedling were investigated. A total of five single-effect QTLs, each for percentages of normal plumules after 4°C treatments for 9 d, 11 d and 14 d, and CSTI and WI, respectively were identified. The QTLs for the percentages of normal plumules after low temperature treatments for 9 d, 11 d and 14 d were on chromosomes 4, 2 and 11, accounting for 14.1%, 17.3% and 21.5% of the phenotypic variation, respectively. QTLs for CSTI and WI were on chromosomes 10 and 1, respectively. Two pairs of epistatic loci were identified, but none of the epistatic loci involved the single-effect QTLs. The RM528-RM340 interval on chromosome 6 interacted with the RM278-RM3919B interval on chromosome 9 for CSTI, and the epistatic interaction accounted for 17.7% of the phenotypic variation. A pair of epistatic loci was identified for WI, the RM246-RM5461 interval on chromosome 1 interacted with the ISA-RM447 interval on chromosome 8, which accounted for 22.6% of the phenotypic variation.

Comparison of physiological and yield traits between purple- and white-pericarp rice using SLs

Five physiological and eleven yield traits of two pairs of sister lines generated from a high generation with similar genetic background (SLs) for purple pericarp were investigated to explore the reasons behind low-yield production of colored rice. Of the five physiological traits examined, except grain anthocyanin content, there were generally similar trends between the P (purple-pericarp) lines and the corresponding W (white-pericarp) lines over two seasons (in the year 2009 and 2010 separately). The results demonstrated that the chlorophyll content of flag leaves, the net photosynthetic rate of flag leaves, and the grain anthocyanin content could be easily influenced by the environment. The physiological functions of the traits for the P lines were more active than those of the corresponding W lines in the year 2010. The grain anthocyanin content of the P lines was much greater in the year 2010 than in the year 2009 during the growth period. The investigation of yield traits revealed that the P lines had reduced 1000-grain weight, yield per plot and grain/brown rice thickness compared to the W lines. A difference comparison of these traits and a source-sink and transportation relationship analysis for these SLs suggested that small sink size was a key reason behind yield reduction of purple pericarp rice.

Pyramiding blast, bacterial blight and brown planthopper resistance genes in rice restorer lines

Abstract Rice blast, bacterial blight (BB) and brown planthopper (BPH) are the three main pests of rice. This study investigated pyramiding genes resistant to blast, BB and BPH to develop restorer lines. Ten new lines with blast, BB and/or BPH resistance genes were developed using marker-assisted selection (MAS) technique and agronomic trait selection (ATS) method. Only HR13 with resistance genes to blast, BB and BPH was obtained. In addition to blast and BB resistance, four lines (HR39, HR41, HR42, HR43) demonstrated moderate resistance to BPH, but MAS for BPH resistance genes were not conducted in developing these four lines. These data suggested that there were unknown elite BPH resistance genes in the Zhongzu 14 donor parent. A more effective defense was demonstrated in the lines with Pi1 and Pi2 genes although the weather in 2012 was favorable to disease incidence. Blast resistance of the lines with a single resistance gene, Pita, was easily influenced by the weather. Overall, the information obtained through pyramiding multiple resistance genes on developing the restorer lines is helpful for rice resistance breeding.

The way to a more precise sheath blight resistance QTL in rice

Rice sheath blight (ShB), caused by Rhizoctonia solani, leads to severe yield losses in many rice production areas worldwide. Resistance to rice ShB is a typical quantitative trait controlled by quantitative trait loci (QTLs). To identify the QTLs responsible for ShB resistance, phenotypes for ShB resistance have been surveyed in different individuals/lines in various mapping populations. The 0–9 rating system, based on the percentage of plant height (PH) above the water line with ShB symptoms, is extensively used in the evaluation of ShB in rice communities. By reviewing previous ShB-QTL-mapping studies, we found that the ShB disease score attained using the 0–9 system was inevitably affected by PH and resulted in the colocalization of QTLs for PH and ShB resistance, i.e., some of the PH-QTLs were mistaken as “ShB-QTLs” and had pleiotropic effects. These colocalizing “ShB-QTLs” are irrelevant to physiological ShB resistance and should not be targeted for utilization. We analyzed the ways in which PH affects ShB resistance through the 0–9 rating system and suggest solutions for improving the accuracy of QTL mapping for rice ShB resistance. We also developed the first physical map of the ShB-QTLs and PH-QTLs detected since 1995. This map will be useful in the marker-assisted selection of ShB-resistant QTLs. Under current circumstances, we believe that conventional breeding remains an effective approach for developing ShB-resistant varieties.

Fine mapping and candidate gene analysis of LM3 , a novel lesion mimic gene in rice

A rice lesion mimic mutant, lm3, was obtained by the mutagenesis of an indica cultivar, 93-11, using γ-ray radiation. Brownish lesions appeared on the leaves of lm3 at the young seedling stage and persisted until the ripening stage. The lm3 mutant was characterised by a shorter plant height and delayed heading compared with the wild-type 93-11. A genetic analysis indicated that the lesion mimic phenotype was controlled by a single recessive gene. Using simple sequence repeat (SSR) markers, the target gene LM3 was first located between marker RM5748 and RM14906 on chromosome 3. We then developed Insertion-Deletion (InDel) markers to fine-map LM3, and the locus was localised to a 29 kb region defined by two InDel markers, In12571 and In12600. Five ORFs were predicted in the candidate region, and DNA sequencing detected a single-nucleotide polymorphism (SNP) in the coding region of LOC Os03g21900. The SNP in the fourth exon (C in 93-11; T in lm3) of LOC_Os03g21900 results in the substitution of a proline (P) with a serine (S) at the 140th amino acid of the deduced uroporphyrinogen decarboxylase protein. We did not detect polymorphisms in the other predicted ORF regions between lm3 and 93-11. These results suggest that LOC_Os03g21900 is the most likely candidate gene for LM3.

Mapping resistant QTLs for rice sheath blight disease with a doubled haploid population

Abstract Sheath blight (SB) disease, caused by Rhizoctonia solani Kuhn, is one of the most serious diseases causing rice (Oryza sativa L.) yield loss worldwide. A doubled haploid (DH) population was constructed from a cross between a japonica variety CJ06 and an indica variety TN1, and to analyze the quantitative trait loci (QTLs) for SB resistance under three different environments (environments 1–3). Two traits were recorded to evaluate the SB resistance, namely lesion height (LH) and disease rating (DR). Based on field evaluation of SB resistance and a genetic map constructed with 214 markers, a total of eight QTLs were identified for LH and eight QTLs for DR under three environments, respectively. The QTLs for LH were anchored on chromosomes 1, 3, 4, 5, 6, and 8, and explained 4.35–17.53% of the phenotypic variation. The SB resistance allele of qHNLH4 from TN1 decreased LH by 3.08 cm, and contributed to 17.53% of the variation at environment 1. The QTL for LH (qHZaLH8) detected on chromosome 8 in environment 2 explained 16.71% of the variation, and the resistance allele from CJ06 reduced LH by 4.4 cm. Eight QTLs for DR were identified on chromosomes 1, 5, 6, 8, 9, 11, and 12 under three conditions with the explained variation from 2.0 to 11.27%. The QTL for DR (qHZaDR8), which explained variation of 11.27%, was located in the same interval as that of qHZaLH8, both QTLs were detected in environment 2. A total of six pairs of digenic epistatic loci for DR were detected in three conditions, but no epistatic locus was observed for LH. In addition, we detected 12 QTLs for plant height (PH) in three environments. None of the PH-QTLs were co-located with the SB-QTLs. The results facilitate our understanding of the genetic basis for SB resistance in rice.

Application of a simplified marker-assisted backcross technique for hybrid breeding in rice

Hybrid rice has contributed greatly to the self-sufficiency of the food supply in China. However, bacterial blight is a major disease that limits hybrid rice production in China. The study was conducted to develop an efficient breeding technique to improve the bacterial blight resistance in hybrid rice. A marker-assisted backcross breeding technique was adopted to improve HN189, an elite restorer line containing the Pi1 gene. This breeding technique was simplified to foreground selection with only one generation of backcrossing and background selection based on phenotypic selection. A novel bacterial blight resistance gene, Xa23, was introgressed into HN189. Two improved restorer lines, HBH145 (with one generation of backcrossing) and HBH146 (with two generations of backcrossing), were obtained that had a significant bacterial blight resistance advantage over HN189. The corresponding hybrid combination Tianyou H145 (Tianfeng A / HBH145) was certified one year earlier than Qianyou H146 (Qianjiang 1A / HBH146). The use of the marker-assisted backcross breeding technique with one generation of backcrossing and without background selection in rice breeding programs shortened the breeding period of the rice.

Combination of Eight Alleles at Four Quantitative Trait Loci Determines Grain Length in Rice.

Grain length is an important quantitative trait in rice (Oryza sativa L.) that influences both grain yield and exterior quality. Although many quantitative trait loci (QTLs) for grain length have been identified, it is still unclear how different alleles from different QTLs regulate grain length coordinately. To explore the mechanisms of QTL combination in the determination of grain length, five mapping populations, including two F2 populations, an F3 population, an F7 recombinant inbred line (RIL) population, and an F8 RIL population, were developed from the cross between the U.S. tropical japonica variety ‘Lemont’ and the Chinese indica variety ‘Yangdao 4’ and grown under different environmental conditions. Four QTLs (qGL-3-1, qGL-3-2, qGL-4, and qGL-7) for grain length were detected using both composite interval mapping and multiple interval mapping methods in the mapping populations. In each locus, there was an allele from one parent that increased grain length and another allele from another parent that decreased it. The eight alleles in the four QTLs were analyzed to determine whether these alleles act additively across loci, and lead to a linear relationship between the predicted breeding value of QTLs and phenotype. Linear regression analysis suggested that the combination of eight alleles determined grain length. Plants carrying more grain length-increasing alleles had longer grain length than those carrying more grain length-decreasing alleles. This trend was consistent in all five mapping populations and demonstrated the regulation of grain length by the four QTLs. Thus, these QTLs are ideal resources for modifying grain length in rice.

Genotype by Environment Interaction: The Greatest Obstacle in Precise Determination of Rice Sheath Blight Resistance in the Field

Rice sheath blight (SB) is the most serious rice disease in China. Resistance of rice to SB is a quantitative trait that is easily influenced by the environment; however, the extent of environmental influence on SB field resistance is still poorly understood. To identify rice genotype by environment interactions for SB resistance, 211 rice genotypes originating from 15 countries were planted and evaluated for SB field resistance in six different environments between 2012 and 2016 after inoculation with the SB pathogen isolate ZJ03. In addition, 65 rice genotypes were evaluated for SB field resistance in another four environments between 2013 and 2016 using ZJ03. Variations in SB field resistance were observed in different genotypes in different environments using objective and subjective rating methods. Two-way analysis of variance indicated that the interaction between the genotype and environment had a highly significant effect on SB field resistance. This analysis indicated that the environment had more of an influence than the genotype itself on SB field resistance, and the genotype by environment interaction was the greatest obstacle in obtaining a precise determination of SB field resistance in rice. The most resistant genotype, GD66, is a good candidate for genetic studies and breeding.