ZmDRR206 functions in maintaining cell wall integrity during maize seedling growth and defense response to external stresses

Plants adaptively change their cell wall composition and structure during their growth,development,and interactions with environmental stresses.Dirigent proteins(DIRs)contribute to environmental adaptations by dynamically reorganizing the cell wall and/or by generating defense compounds.A maize DIR,ZmDRR206,was previously reported to play a dominant role in regulation of storage nutrient accumulation in endosperm during maize kernel development.Here we show that ZmDRR206 mediates maize seedling growth and disease resistance by coordinately regulating biosynthesis of cell wall components for cell-wall integrity(CWI)maintenance.Expression of ZmDRR206 was induced in maize seedlings upon pathogen infection.ZmDRR206 overexpression in maize resulted in reduced seedling growth and photosynthetic activity but increased disease resistance and drought tolerance,revealing a tradeoff between growth and defense.Consistently,ZmDRR206 overexpression reduced the contents of primary metabolites and down-regulated genes involved in photosynthesis,while increasing the contents of major cell wall components,defense phytohormones,and defense metabolites,and up-regulated genes involved in defense and cell-wall biosynthesis in seedlings.ZmDRR206-overexpressing seedlings were resistant to cell-wall stress imposed by isoxaben,and ZmDRR206 physically interacted with ZmCesA10,which is a cellulose synthase unit.Our findings suggest a mechanism by which ZmDRR206 coordinately regulates biosynthesis of cell-wall components for CWI maintenance during maize seedling growth,and might be exploited for breeding strong disease resistance in maize.

Genetic dissection of yield-related traits in response to drought stress in common bean

Drought stress severely impairs common bean production.For facilitating drought-resistance breeding in common bean,molecular markers were identified in a genome-wide level marker–trait association study.A panel of 210 common bean accessions showed large variation in 11 agronomic traits at the adult stage(plant height,pod number per plant,seed number per pod,seed number per plant,seed yield per plant,pod length,harvest index,pod harvest index,days to maturity,hundred-seed weight,and seed yield)under two water conditions.The coefficient of variation ranged from 6.21% for pod harvest index to51.00% for seed number per plant under well-watered conditions,and from 4.05% for days to maturity to 40.72% for seed number per plant under drought stress.In a genome-wide association study,119quantitative-trait loci were associated with drought resistance,including 41 adjacent to known loci.Among these loci,12 were found to be associated with at least two traits.Three major loci were identified at Pv01 and Pv02.A set of candidate genes were found that encode MYBs,AREBs,WKRYs,and protein kinases.These results reveal promising alleles that control drought resistance,shedding light on the genetic basis of drought resistance and accelerating future efforts for drought resistance improvement in common bean.

Rational Design of Grain Size to Improve Rice Yield and Quality

Grain size, determined by grain length, grain width and grain thickness, is associated with grain yield and quality. Many genes controlling grain size were cloned and their related regulatory mechanisms were clearly clarified. However, whether these genes can be directly introduced into japonica rice for grain size improvement is unknown.

Construction of a high-density SSR genetic linkage map and identification of QTL for storage-root yield and dry-matter content in sweetpotato

Sweetpotato(Ipomoea batatas(L.)Lam.)is a widely grown food crop especially in developing countries.Increasing storage-root yield and dry-matter content has been the main breeding objective of the crop,and DNA marker-assisted breeding is needed for this purpose.In this study,using a mapping population of 500 F1 individuals from a cross between Xushu 18(female)and Xu 781(male),we constructed a highdensity genetic linkage map of sweetpotato using 601 simple-sequence repeat(SSR)primer pairs.The Xushu 18 map contained 90 linkage groups with 5547 SSR markers and spanned 18,263.5 cM,and the Xu 781 map contained 90 linkage groups with 4599 SSR markers and spanned 18,043.7 cM,representing the highest genome coverage yet reported for sweetpotato.We identified 33 QTL for storage-root yield and 16 QTL for dry-matter content,explaining respectively 6.5%–47.5%and 3.2%–18.9%of variation.These results provide a foundation for fine-mapping and cloning of QTL and for marker-assisted breeding in sweetpotato.

Additive Effects of QTLs/Genes on Rice Grain Size Traits Revealed by Genetic Comparisons

Grain size is controlled by many QTLs and/or genes.Despite intensive study,the genetic interactions between these QTLs/genes remain largely unclear.We previously found that the japonica large-grain rice cultivar TD70 with 1000-grain weight of 68.6 g carries superior alleles at the loci GW2,GS3,qGL3,GS5 and GW8,all of which are known to control grain size,while the indica rice variety Kasalath with 1000-grain weight of 19.1 g has normal alleles at these loci.

Efficient Improvement of Nutritional Content in Rice Grains by Precise Base Editing of OsROS1

Aleurone forms the outermost layer of the rice endosperm and plays a critical role in apoplastic nutrient uptake during endosperm development.Thickening the aleurone layer has been proposed to significantly increase the nutrient content of rice grains.In this study,we used a CRISPR/Cas9-mediated precise base editing method to target OsROS1,a gene associated with aleurone thickness,in the background of the japonica glutinous rice cultivar Zhennuo 19。

A Cleaved Amplified Polymorphic Sequence Marker to Detect Variation in Wx Locus Conditioning Translucent Endosperm in Rice

The translucent endosperm trait in a japonica rice variety ＇Kantou 194＇ is controlled by a Wx-mq gene which is allelic to Wx locus by genetic analysis and allelic test. The amylose content analysis showed that an intermediate amylose content between those of glutinous and non-glutinous rice existed in endosperm of homozygous Wx-mq genotype. The slight changes of amylose content in different varieties and F1 grains with an identical Wx-mq genotype might be influenced by dissimilar genetic background. To identify the Wx-mq genotype simply and rapidly, a cleaved amplified polymorphic sequence （CAPS） marker was designed. The result from the molecular detection indicated that it could be used for marker-assisted selection for low amylose content varieties in rice breeding.

Molecular diversity and genetic structure of 380 sweetpotato accessions as revealed by SSR markers

Sweetpotato, Ipomoea batatas（L.） Lam., is an important food crop widely cultivated in the world. Evaluation of genetic relationships among diverse cultivars and landraces is necessary for efficient exploitation of genetic diversity in the existing germplasm resources. In the present study, a collection of 380 sweetpotato accessions assembled from different agro-climatic zones of China and other countries were genotyped using 30 SSR primer pairs. Model-based structure analysis separated the germplasm into three populations, P1, P2 and P3, containing 228, 133 and 19 accessions, respectively, which was consistent with the results of phylogenic and principal component analysis（PCA）. Analysis of molecular variance（AMOVA） revealed significant genetic differentiation among inferred populations, accounting for 16.47% of the total molecular variance, however, the differences between the regions were not significant, the total variation were due to the differences between the genotypes within the population. Pairwise fixation index（F ST） suggested that populations P1 and P3 had the highest differentiation, while populations P1 and P2 had the lowest differentiation. The diversity among populations was wide, which confirmed the genetic distinction of populations. Through comparing model-based structure and domestication-based classification, it was found that the accessions of population P1 mainly belonged to modern cultivars, and the accessions of populations P2 and P3 basically corresponded to landraces, by which we suggest that modern cultivars maybe had experienced a two-step domestication history. Our results illustrated clear genetic relationships among 380 sweetpotato accessions, exhibiting the potential of accelerating the process of future sweetpotato breeding program by molecular marker based parental selection.

Stability of QTL Across Environments and QTL-by-Environment Interactions for Plant and Ear Height in Maize

Better understanding of genotype-by-environment interaction （GEI） is expected to provide a solid foundation for genetic improvement of crop productivity especially under drought-prone environments. To elucidate the genetic basis of the plant and ear height, 2 F2：3 populations were derived from the crosses of Qi 319 × Huangzaosi （Q/H） and Ye 478 × Huangzaosi （Y/H） with 230 and 235 families, respectively, and their parents were evaluated under 3 diverse environments in Henan, Beijing, and Xinjiang, China during the year of 2007 and 2008, and all the lines were also evaluated under water stress environment. The mapping results showed that a total of 21 and 12 QTLs were identified for plant height in the Q/H and Y/H population, respectively, and 24 and 13 QTLs for ear height, respectively. About 56 and 73% of the QTLs for 2 traits did not present significant QTL-by-environment interaction （QE1） in the normal joint analyses for Q/H and Y/H population, respectively, and about 73% of the QTLs detected did not show significant QEI according to joint analyses for stress condition in Q/H. Most of the detected major QTLs exhibited high stability across different environments. Besides, several major QTLs were detected with large and consistent effect under normal condition （Chr. 6 and 7 in Q/H; Chr. 1, 3 and 9 in Y/H）, or across 2 water regimes （Chr. 1, 8 and 10 for in Q/H）. There were several constitutive QTLs （3 for Q/H and 1 for Y/H） with no or minor QTL-by-environment for the 2 populations. Finally, we found several genomic regions （Chr. 1, 10, etc.） to be co-located across the populations, which could provide useful reference for genetic improvement of these traits in maize breeding programs. Comparative genomic analysis revealed that 3 genes/genetic segments associated with plant height in rice were orthologous to these 3 identified genomic regions carrying the major QTLs for plant and ear height on Chr. 1, 6, and 8, respectively.

Characterization of eating quality and starch properties of two Wx alleles japonica rice cultivars under different nitrogen treatments

To understand the effect of nitrogen(N) fertilizer on rice(Oryza sativa L.) eating and cooking quality(ECQ). Here, we investigated the ECQ attributes, physicochemical foundation of ECQ, and amylopectin fine structure of two Waxy(Wx) alleles japonica rice cultivars Nanjing 9108(NJ9108) and Huaidao 5(HD5) under four N rates(0, 150, 300, and 450 kg ha^-1). Sensory and pasting properties of the two cultivars varied depending on N rates. Compared with the control(0 kg ha^-1), the overall eating quality and sensory value were significantly decreased under the N rates of 300 and 450 kg ha^-1. Further, conventional descriptive analysis showed that the stickiness and retrogradation of cooked rice were significantly decreased. These results indicated that application of N fertilizer seems to affect the texture of cooked rice, causing it to be less sticky, lowering its retrogradation, and consequently reducing its palatability. Results from rapid visco analyzer(RVA) revealed that the peak and breakdown viscosities significantly decreased, while the setback viscosity and peak time increased under the N rates of 300 and 450 kg ha^-1. However, no significant difference was observed when the N rate was 150 kg ha^-1, indicating that less N fertilization can maintain rice ECQ. As the N rate increasing, protein content increased, whereas apparent amylose content, starch content, and gel consistency almost unchanged. Interestingly, compared with the control, under N treatments, the percentage of short amylopectin branches in NJ9108 was decreased, but increased in HD5, as controlled by amylopectin synthesis-related genes. Notably, SSI and BEIIb were down-regulated in NJ9108, whereas BEIIb was up-regulated in HD5. Thus, the palatability of both rice cultivars was significantly decreased under excessive N fertilization as a consequence of reduced stickiness and retrogradation of the cooked rice, which might have resulted from an elevated protein content and altered amylopectin fine structure. In addition, amylopectin synthesis appeared to be affected by N fertilizer and the genotype of the rice cultivar.

Research progress on the breeding of japonica super rice varieties in Jiangsu Province,China

In this study we report the results of a decade-long breeding program for japonica super rice made by Nanjing Branch of Chinese National Center for Rice Improvement in Jiangsu Academy of Agricultural Sciences. We concluded that selection of parents with good comprehensive traits and complementary advantages and disadvantages of both parents in the hybrid combination, and early selection of high heritability traits in earlier segregating generations could significantly improve the breeding efficiency. The use of closely-linked functional markers in pyramiding of multiple genes could greatly increase breeding efficiency, avoiding time-consuming and laborious steps that were used in traditional breeding program. It is also important to coordinate the yield components with variety characteristics such as yield stability, wide adaptability, lodging resistance, and an attractive grain appearance during late growth stage of rice.

Production of Two Elite Glutinous Rice Varieties by Editing Wx Gene

The waxy gene(Wx) in rice, which encodes the granule bound starch synthase enzyme, is responsible for amylose synthesis. Glutinous(sticky) rice has little or no amylose that can be used in various applications, such as brewing. In this study, knockout of the Wx gene with CRISPR/Cas9 technology was conducted in two elite japonica rice lines, Huaidao 5(HD5) and Suken 118(SK118), aiming to develop elite sticky rice varieties. We achieved six homozygous T_0 plants with more than 200 bp deletion in the Wx gene, as well as 36 wx-HD5 and 18 wx-SK118 homozygous transgene-free plants in the T_1 generation. The seeds of all the mutants were white and opaque, similar to those of sticky rice, and contained only 2.6%–3.2% amylose. Results of scanning electron microscopy showed that the quality of rice did not change. In conclusion, we successfully developed two elite sticky rice varieties.

Overexpression of IbPAL1 promotes chlorogenic acid biosynthesis in sweetpotato

Sweetpotato[Ipomoea batatas(L.)Lam.],a food crop with both nutritional and medicinal uses,plays essential roles in food security and health-promoting.Chlorogenic acid(CGA),a polyphenol displaying several bioactivities,is distributed in all edible parts of sweetpotato.However,little is known about the specific metabolism of CGA in sweetpotato.In this study,IbPAL1,which encodes an endoplasmic reticulum-localized phenylalanine ammonia lyase(PAL),was isolated and characterized in sweetpotato.CGA accumulation was positively associated with the expression pattern of IbPAL1 in a tissue-specific manner,as further demonstrated by overexpression of IbPAL1.Overexpression of IbPAL1 promoted CGA accumulation and biosynthetic pathway genes expression in leaves,stimulated secondary xylem cell expansion in stems,and inhibited storage root formation.Our results support a potential role for IbPAL1 in sweetpotato CGA biosynthesis and establish a theoretical foundation for detailed mechanism research and nutrient improvement in sweetpotato breeding programs.

QTL Mapping for Grain Size Traits Based on Extra-Large Grain Rice Line TD70

Grain size traits, including grain length, grain width and grain thickness, are controlled by quantitative trait loci （QTLs）. Many QTLs relating to rice grain size traits had been reported, but their control mechanisms have not yet been elucidated. A recombinant inbred line （RIL） population of 240 lines, deriving from a cross between TD70, an extra-large grain size japonica line with 80 g of 1000-grain weight, and Kasalath, a small grain size indica variety, were constructed and used to map grain size QTLs to a linkage map by using 141 SSR markers in 2010 and 2011. Five QTLs for grain length, six for grain width and seven for grain thickness were detected distributing over chromosomes 2, 3, 5, 7, 9 and 12. Seven QTLs, namely qGL3.1, qGW2, qGW2.2, qGW5.1, qGW5.2, qGT2.3 and qGT3.1, were detected in either of the two years and explained for 56.19%, 4.42%, 29.41%, 10.37%, 7.61%, 21.19% and 17.06% of the observed phenotypic variances on average, respectively. The marker interval RM1347-RM5699 on chromosome 2 was found common for grain length, grain width and grain thickness; qGL3.1 and qGT3.1 were mapped to the same interval RM6080-RM6832 on chromosome 3. All 18 QTL alleles were derived from the large grain parent TD70. Most of the QTLs mapped in the present study were found the same as the genes previously cloned （GW2, GS3 or qGL3, GW5 and GS5）, and several were the same as the QTLs （GS7 and qGL-7） previously mapped. Three QTLs, qGL2.2 on chromosome 2, qGW9 and qGT9 on chromosome 9, were first detected. These results laid a foundation for further fine mapping or cloning of these QTLs.

Inheritance Analysis and QTL Mapping of Rice Starch Viscosity (Rapid Visco Analyzer Profile) Characteristics

The rice starch viscosity characteristics, which can be indicated by Rapid Visco Analyzer profile （RVA profile）, have been proved useful for the evaluation of cooking and eating quality in rice breeding program. To study the inheritance of the RVA profile, an F2 population of Wuyujing 3/Aichi 106 was used. The results indicated that the peak viscosity （PKV） was a typical quantitative character, and the hot paste viscosity （HPV）, cool paste viscosity （CPV）, setback viscosity （SBV）, breakdown viscosity （BDV）, and consistence viscosity （CSV） might be controlled by one major gene and several minor genes To elucidate the genetic basis of the paste viscosity characteristics, a recombinant inbred line （RIL） population derived from Nikken 2/Milyang 23 and its genetic linkage map were used to map the QTLs controlling RVA profiles in 2005 and 2006. A total of 34 QTLs distributed on chromosomes 1,2, 3, 4, 6, 7 and 8 were detected, including 19 and 15 QTLs in 2005 and 2006 respectively. Eight QTLs were both detected in the two years, qHPV6, qCPV6, qCSV6, qSBV6, and qBDV6 were located on chromosome 6, while qHPV2, qCSV2, and qCPV2were on chromosome 2.

Cluster Analysis on Japonica Rice(Oryza sativa L.) with Good Eating Quality Based on SSR Markers and Phenotypic Traits

Diversity of 60 conventional japonica rice accessions with good eating quality at home and abroad was analyzed using SSR molecular markers, agronomic traits and taste characteristics. A total of 290 alleles were detected in the 60 accessions at 72 SSR loci with the high similarity coefficients varying between 0.600 and 0.924. The loci on chromosome 5 showed the greatest value in average allele number. Additionally, most of the SSR loci could detect 3 to 4 alleles. An UPGMA dendrogram based on the cluster analysis of the genetic similarity coefficients showed that the grouping trend of part of the rice accessions was geographic-related and most of the rice accessions in Jiangsu Province, China were clustered together. Furthermore, many domestic accessions from south and north origins in China were close to the foreign japonica rice varieties, as proved by their pedigree origin from the foreign high-quality sources. For taste characteristics, part of the accessions with excellent taste were clearly clustered into one category though they came from different geographical regions, which indicates that taste characteristics of some varieties were mainly genetically determined. In addition, the agronomic traits of japonica rice with good taste might be closely related with their geographical origins, but the relationship between superior taste characteristics and agronomic traits should be further clarified.

Plant Temperature and Its Simulation Model of Thermo-Sensitive Genic Male Sterile Rice

Plant temperature （Tp） and its relations to the microclimate of rice colony and irrigation water were studied using a thermo-sensitive genic male sterile （TGMS） rice line, Pei＇ai 64S. Significant differences in the daily change of temperature were detected between Tp and air temperature at the height of 150 cm （TA）. From 8：00 to 20：00, Tp was lower than TA, but they were similar during 21：00 to next 7：00. The maximum Tp occurred one hour earlier than the maximum TA, though they both reached the minimum at 6：00. Tp fluctuated less than TA. At the same height, during 6：00-13：00, Tp was higher than air temperature （Ta）, and Tp reached the maximum one hour earlier than Ta. During the rest time on sunny day, Tp was close to or even a little lower than Ta. On overcast day, Tp was higher than Ta in the whole day, and both maximized at the same time. In addition, Tp was regulated by solar radiation, cloudage and wind speed in daytime, and by irrigation water at night. The present study indicated that a TA of 29.6℃ was the critical point, at which Tp was increased or decreased by irrigation water. Tp and the difference between water and air temperatures showed a conic relation. Tp fluctuation was also regulated by the absorption or reflection of solar radiation by leaves during daytime and release of heat energy during nighttime. By analysis on correlation and regression simulation, two models of Tp were established.

The Comparison in Tissue Culture Ability of Mature Embryo in Different Cultivars of Rice

In order to study the regeneration technology of mature embryos in different rice varieties,nine japonica,nine indica and eleven hybrid rice varieties of two line or three line or superiority combinations were selected as explants to study the callus induction,differentiation and regeneration rates on different media.The higher callus induction （61.7-89.2%） was observed in japonica rice,when cytokinin was added at lower concentration （0.3 mg L-1 6-BA） in M8 basal medium,supplemented with 30 g L-1 sucrose,8 g L-1 agar and 2 mg L-1 2,4-D.Further,the addition of two cytokinins （2 mg L-1 6-BA,0.5 mg L-1 KT） and 1 mg L-1 NAA in the M8 basal supplemented medium resulted in 9.1-100% of the callus induction in indica rice.The percent callus induction in hybrid rice varieties was 40-86.3% when addition of 1 mg L-1 6-BA and 1 mg L-1 KT was added,and the cytokinins was required by the japonica and indica rice varieties in the M8 basal supplemented medium.It was observed that when the 0.5 mg L-1 2,4-D and 1 mg L-1 6-BA were added in japonica rice,and 0.2 mg L-1 2,4-D and 0.5 mg L-1 6-BA were added in indica and hybrid rice in the MS different media,the regeneration rates were 9.2-59.5%,3.6-87.5% and 17.2-43.2% for japonica,indica and hybrid rice,respectively.Thus,the regeneration technology with higher output is established in the mature embryos of similar rice varieties.

Genetic Improvement of Japonica Rice Variety Wuyujing 3 for Stripe Disease Resistance and Eating Quality by Pyramiding Stv-b^i and Wx-mq

Japonica rice variety Kanto 194 as the donor of resistance gene Stv-b^i and Iow-amylose content gene Wx-mq was used to improve the resistance and eating quality of Wuyujing 3 by the breeding strategy of backcross. In continuous backcross and selfcross generations, the related molecular markers with Stvobi and Wx-mq genes were utilized for genotypic detection by associated with resistance identification of rice stripe disease and agronomic traits selection. Finally, 10 improved lines with homozygous genotype Stv-b^i Stv-b^iWx-mqWx-mq were obtained from BC3F4 generation. The results of comparative analysis indicated that most characters of these lines were consistent with those of recipient parent Wuyujing 3, except for the improved resistance, appearance and eating quality. By evaluation of the comprehensive performance of them, two excellent lines K01 and K04 were selected for further experiments.