Construction of Near Isogenic Lines for Pericarp Color and Evaluation on Their Near isogenicity in Rice

The inheritance of purple pericarp was studied through genetic analysis using F2, BC1F1 and F3 from a cross between a purple pericarp rice variety Zixiangnuo and a white pericarp rice variety Chunjiangnuo 2. Seven pairs of near isogenic lines （NILs） for pericarp color of rice were further constructed. Genetic analysis indicated that the purple pericarp was controlled by two complementary genes （Pb and Pp）. Agronomic trait analysis and polymorphism analysis using SSR markers on these seven pairs of NILs were used in the evaluation of near-isogenicity. No significant differences in agronomic traits were found except 1000-grain weight between the NILs. The polymorphic SSR markers for the parents were only detected in target segments of the five pairs of NILs different in Pb, which revealed the ideal near isogenicity of them. However, for the two pairs of NILs different in Pp, the polymorphic markers for the two parents were detected as well as in non-targeted segments of chromosomes 11 and 12, respectively.

Integrating transcriptomic and proteomic analyses of photoperiod-sensitive in near isogenic maize line under long-day conditions

As a short-day plant species, maize requires an optimal photoperiod for inducing reproductive growth. However, there is a lack of information regarding photoperiod-induced changes in maize mRNA and protein levels. In this study, a photoperiod-insensitive maize inbred line and its near isogenic photoperiod-sensitive line were used. By integrating RNAbased transcriptomic and iTRAQ LC-MS/MS-based proteomic approaches, we generated a comprehensive inventory of the transcripts and proteins with altered abundances in response to a long photoperiod(LP) during growth stage transitions. We detected 22 000 transcripts in RNA-sequence runs and 5 259 proteins from an iTRAQ-based analysis. A weak correlation between mRNA-and protein-level changes was observed, suggesting the LP-induced transition between maize growth stages is largely regulated post-transcriptionally. Differentially expressed genes influenced by LP conditions were associated with several regulatory processes in both maize inbred lines, especially phosphate ion transport and the circadian rhythm. Additionally, 31 transcripts and six proteins related to photoperiodic flowering in maize were identified by comparing transcriptomic and proteomic data. This transcriptomic and proteomic analysis represents the first comprehensive and comparative study of gene/protein-level changes occurring in photoperiod-sensitive and-insensitive maize inbred lines during growth stage transitions under LP conditions.

Development of branchless watermelon near isogenic lines by marker assisted selection

Pruning is time-consuming and laborious in watermelon cultivation,which can not meet the needs for simplified cultivation in the future.The development of branchless lines will provide important germplasms for breeding watermelon varieties and is an important method for genetic improvement.In this study,the watermelon accession,Wu Cha Zao(WCZ)is a branchless inbred line that carries the branchless gene Clbl,which was used as the donor parent to develop branchless near isogenic lines(NILs).To construct the NILs of Clbl,WCZ crossed with the normal branching watermelon inbred line WT20 which was used as the recurrent parent.The co-segregating markers dCAPS10 and Indel1 with Clbl were used for foreground selection,and a total of 108 SSR markers was selected with good polymorphism between two parental lines for background selection which had relatively uniform distribution across 11 chromosomes.Using these markers to select individuals from the BC_(1)F_(1),BC_(2)F_(1),and BC_(2)F_(2) generations,three NILs with a proportion of recurrent parent genome(PRPG)>99%were finally obtained.The lateral branch and plant height phenotypes did not significantly differ between the NILs and WCZ,indicating that the NILs of Clbl under the genetic background of WT20 has been successfully developed.These results provide ideal materials for further in-depth analysis of the genetic mechanisms of lateral branch development and ideal plant architecture breeding in watermelon.

Dissection of QTLs for Yield Traits Using Near Isogenic Lines Derived from Residual Heterozygous Lines in Rice

Three residual heterozygous lines (RHLs) carrying heterozygous segments in the intervals RM587–RM225, RM204–RM6119 and RM6119–RM402 on the short arm of rice chromosome 6, respectively, were selected from a rice population derived from an RHL for the interval RM587–RM402. Ten maternal homozygotes, 10 paternal homozygotes and 20 heterozygotes were selected from each of the F2 populations derived from the three RHLs. The three sets of near isogenic lines (NILs) were grown to detect the grain yield per plant, number of panicles per plant, number of filled grains per panicle and 1000-grain weight. With analysis on the phenotypic differences among the three genotype groups in each NIL set and those among overlapping chromosome segment substitution lines, three QTLs for number of filled grains per panicle and two QTLs for grain yield per plant were resolved. They were located in the intervals ranging from 0.66 Mb to 2.49 Mb. The additive effect was higher than the dominance effect at each locus. The allele for increasing the trait value was derived from the paternal parent at qNFGP6-1, and from the maternal alleles at other QTLs. Based on the present study, an approach for constructing new genetic resource to facilitate fine mapping of QTLs in rice was proposed.

Identification of AFLP Markers Linked to Leaf Rust Resistance Genes Using Near Isogenic Lines of Wheat

The present investigation was undertaken to find molecular markers linked to leaf rust resistance genes, Lr9 and Kharchia local mutant KLM4-3B. Preliminary AFLP analysis was carried out with different stocks, a survey of primer combinations with different selective nucleotide indicated that for each primer combination, the number of scorable loci ranged from 34 to 123. Only a limited primer combination used in the set of parental and near isogenic lines showed a high level of polymorphism for AFLP marker. Putative AFLP marker were found to be linked to Lr9, Lr19 and KLM4-3B. The alien genes were readily identified.

Cottonseed Protein, Oil, and Mineral Nutrition in Near-Isogenic <i>Gossypium hirsutum</i>Cotton Lines Expressing Leaf Color Phenotypes under Field Conditions

Information about the effects of phenotype traits on cottonseed protein, oil, and nutrients is scarce. The objective of this research was to investigate the effects of leaf color trait on seed nutrition in near-isogenic Gossypium hirsutum cotton expressing green (G) and yellow (Y) leaf color phenotypes. Our hypothesis was that leaf color can influence the accumulation of nutrients in seeds. Sets of isogenic lines were: DES 119 (G) and DES 119 (Y);DP 5690 (G) and DP 5690 (Y);MD 51ne (G) and MD 51ne (Y);SG 747 (G) and SG 747 (Y). Each NIL set is 98.44 % identical. Parent line SA 30 (P) was used as the control. The experiment was repeated for two years (2014 and 2015). The results showed that, in 2014, seed oil in DES 119 (G) and SG 747 (G) were significantly higher than their equivalent yellow lines. Green lines showed higher content of phosphorus compared with yellow lines. Higher levels of Cu, Fe, Mn, Ni, and Zn were recorded in DES 119 (G) and MD 51ne (G). In 2015, seed protein, oil, C, N, P, B, Cu, and Fe were higher in green lines than in yellow lines. There was a significant correlation between protein and nutrients, and between oil and nutrients in 2015, but not in 2014 as the temperature was warmer in 2015 than in 2014. This research demonstrated that leaf color can alter seed composition and mineral nutrition under certain environmental growing conditions such as temperature.

Identification of Molecular Markers Linked to Leaf Rust Resistance Genes in Wheat and Their Detection in the Local Near-Isogenic Line

Sixty-five random amplified polymorphic DNA (RAPD) primers were used for the detection of polymorphism among recipient and donor parents and their isogenic lines linked to leaf rust resistance genes, Lr9 and the resistant gene in kharchia local mutant KLM4-3B. Three primers showed polymorphism among recurrent parent, donor parent and isogenic lines.

Blue-White Colony Selection of Virus-Infected Isogenic Recipients Based on a Chrysovirus Isolated from Penicillium italicum

Mycoviruses have been found to infect more than 12 species of Penicillium,but have not been isolated from Penicillium italicum(P.italicum).In this study,we isolated and characterized a new double-stranded RNA(dsRNA)virus,designated Penicillium italicum chrysovirus 1(PiCVl),from the citrus pathogen P.italicum HSPi-YNl.Viral genome sequencing and molecular characterization indicated that PiCV 1 was highly homologous to the previously described Penicillium chrysogenum virus.We further constructed the mutant HSPi-YNl ApksP defective in the polyketide synthase gene(pksP),which is involved in pigment biosynthesis,and these mutants formed albino(white)colonies.Then we applied hyphal anastomosis method to horizontally transmit PiCV 1 from the white virus-donors(i.e.,HSPi-YNl mutants)to wild-type recipients(i.e.,P.italicum strains HSPi-CQ54,HSPi-HB4,and HSPi-HNl),and the desirable PiCV 1-infected isogenic recipients,a certain part of blue wild-type strains,can be eventually selected and confirmed by viral genomic dsRNA profile analysis.This bluewhite colony screening would be an easier method to select virus-infected P.italicum recipients,according to distinguishable color phenotypes between blue virus-recipients and white virus-donors.In summary,the current work newly isolated and characterized PiCVl,verified its horizontal transmission among dually cultured P.italicum isolates,and based on these,established an effective and simplified approach to screen PiCV 1-infected isogenic recipients.

Fine Mapping of qTGW3-1,a QTL for 1000-Grain Weight on Chromosome 3 in Rice

The QTL qTGW3-1 was located on chromosome 3 of rice （Oryza sativa L.） and associated with the 1 000-grain weight （TGW） according to the result of our earlier study. With the objective of fine mapping of this locus, we developed a F2 population consisting of 3 428 plants derived from the cross between TGW-related near isogenic line DL017 （BC3F4 generation of GSL 156×Nipponbare） and the recurrent parent Nipponbare. Using six microsatellites, this QTL was delimited between RM5477 and RM6417. Markers MM 1455 and MM 1456 within this region were used for further mapping of this QTL. Finally, qTGW3-1 was fine-mapped into a 89-kb interval between RM5477 and MM1456, which locates in the BAC clone AC107226 harboring five putative candidate genes.

Photosynthesis and Dry Matter Accumulation in Different Chlorophyll-Deficient Rice Lines

Three different chlorophyll-deficient rice isogenic lines chl,fgl and pgl,and their recurrent parent zhefu802 （zf802） were used to study effects of leaf color on photosynthesis,dry matter accumulation,yield,and quality in early season indica rice.Analysis showed that the chlorophyll （Chl.） a/b ratio of isogenic lines chl-8,pgl and fgl was 5.35,10.00 and 15.46,respectively,among them,line fgl had higher leaf area index （LAI）,higher net photosynthetic rate and higher grain-filling rate than its recurrent parent zf802 at the later period of grain filling stage;while LAI,net photosynthetic rate and dry matter accumulated in lines chl-8 and pgl were lower than in zf802.Differences were found in the grain yield and quality among chlorophyll deficient isogenic lines,lines fgl,chl-8 and zf802 had similar grain yield,which was significantly higher than that of pgl;the highest milling quality was observed in isogenic line fgl,with relatively high protein content.This study showed that isogenic line fgl would become a unique material for the development of high yield rice with high grain quality because of its slow aging process and relative steady grain-filling rate.

Fine Mapping and Candidate Gene Analysis of Resistance Gene R_(SC3Q) to Soybean mosaic virus in Qihuang 1

Soybean mosaic virus （SMV） disease is one of the most destructive viral diseases in soybean （Glycine max （L.） Merr.）. SMV strain SC3 is the major prevalent strain in huang-huai and Yangtze valleys, China. The soybean cultivar Qihuang 1 is of a rich resistance spectrum and has a wide range of application in breeding programs in China. In this study, F1, F2 and F2：3 from Qihuang 1×nannong 1138-2 were used to study inheritance and linkage mapping of the SC3 resistance gene in Qihuang 1. The secondary F2 population and near isogenic lines （nILs） derived from residual heterozygous lines （RhLs） of Qihuang 1×nannong 1138-2 were separatively used in the ifne mapping and candidate gene analysis of the resistance gene. Results indicated that a single dominant gene （designated RSC3Q） controls resistance, which was located on chromosome 13. Two genomic-simple sequence repeat （SSR） markers BARCSOYSSR_13_1114 and BARCSOYSSR_13_1136 were found lfanking the two sides of the RSC3Q. The interval between the two markers was 651 kb. Quantitative real-time PCR analysis of the candidate genes showed that ifve genes （Glyma13g25730, 25750, 25950, 25970 and 26000） were likely involved in soybean SMV resistance. These results would have utility in cloning of RSC3Q resistance candidate gene and marker-assisted selection （MaS） in resistance breeding to SMV.

Development of Heat Tolerant Two-Line Hybrid Rice Restorer Line Carrying Dominant Locus of OsHTAS

In order to create novel germplasm resources for breeding heat tolerant variety,we transferred a dominant allele OsHTAS,previously characterized and cloned from a high-temperature tolerant local variety HT54,which was collected from the rice production area of southern China,into a high-temperature sensitive intermediate breeding line HT13 through six rounds of successive backcross by using marker-assisted selection.The molecular analysis showed that the recovery of genetic background of a resultant near isogenic line(NIL),MHT13,was around 99.8%.The OsHTAS gene introduced in the MHT13 expressed normally in the HT13 genetic background,mediating heat tolerance and phenotype similar to those of the donor parent HT54.The major agronomic traits of MHT13 resembled those of the recurrent parent HT13.Moreover,MHT13 had high general combining ability and its rice quality reached the grade 3 standard of edible high-quality rice issued by Ministry of Agriculture of the People’s Republic of China,which greatly improved its application value in rice production.

Effect of Dominant Semi-Dwarf Gene on Plant Height and Its Related Traits and Sensitivity to Gibberellic Acid in Rice

Six pairs of tall and dwarf near-isogenic lines derived from a dominant semi-dwarf mutant （Y98149） were selected to study height expression and sensitivity to gibberellic acid （GA3）. The lengths of the 4-5th internode, the 3rd, 2nd, 1st internodes from the top and the panicle length in the six dwarf near isogenic lines were 97.2%, 53.3%, 65.1%, 61.9% and 94.7% of those in the six tall ones, respectively, indicating that the dominant semi-dwarfing gene significantly inhibited the internode elongation. Moreover, Y98149 （mutant type） was more sensitive to GA3 than Y98148 （wild type）, and had a lower GA3 concentration in plant, about 78% of Y98148.

Identification of loci and candidate genes controlling kernel weight in barley based on a population for which whole genome assemblies are available for both parents

Kernel weight(KW), together with kernel number per unit area, determines yield of cereal crops. Here,two barley recombinant inbred lines(RILs) populations with a shared parent were used to identify loci controlling KW. One is Baudin/AWCS276(BA) for which a linkage map was available. Several largeeffect QTL controlling KW were detected in this population. Another is Morex/AWCS276(MA). A linkage map with 5273 makers formed 1454 clusters, was constructed by the genotyping by sequence(GBS) data of 201 RILs from this population. A single marker was selected to represent each of the clusters and the linkage map constructed with these markers covers a total length of 1022.4 c M with an average interval of approximately 0.7 cM between loci. Three of the large-effect loci controlling KW(located on 2 HL, 6 HL,and 7 HL, respectively) identified from the BA population were also detected in the MA population under different environments. The locus on 6 HL was detected in each of the experiments conducted for both populations thus was selected for developing near isogenic lines(NILs). Apart from KW, the two isolines for each pair of the putative NILs obtained showed no significant difference for any of the morphological characteristics assessed. The average difference in KW between the isolines for all the NILs obtained was about 15% based on assessments under both glasshouse and field conditions. Taken advantage that high quality genome assemblies for both Morex and AWCS276 are available, we identified candidate genes underlying two of the three loci based on an orthologous analysis. The NILs developed and the candidate genes identified in this study should facilitate the cloning and functional analysis of genes regulating KW in barley.

Genetic Identification of a New Small Grain Dwarf Gene in Rice

The plant material used in the study was rice line 162d, a new small grain dwarf mutant. Polymorphic analysis of 221 SSR loci demonstrated that 162d derived from a semidwarf variety Shuhui 162 through mutation, and 162d and Shuhui 162 were just a pair of near isogenic lines. Genetic analysis of F_1 and F_2 populations suggested that dwarfism in 162d was controlled by a single recessive gene. Phenotypic characteristics of the mutant gene were that plant height was about a quarter of normal height, grain size about a quarter of normal size, leaf was short and broad, and seed setting rate was very low, compared with the near isogenic line Shuhui 162. The mutant gene was sensitive to gibberellin (GA_3) treatment and did not located on the region near the centromere of rice chromosome 5, where dl gene located. Therefore, it was concluded that the mutant gene of 162d was a new small grain dwarf gene in rice.

Performance of Conventional and Transgenic Soybean Cultivars in the South and Alto Paranaiba Regions of Minas Gerais, Brazil

The aim of this study was to compare transgenic and conventional soybean cultivars in regard to yield performance and agronomic traits. Six soybean cultivars were tested: three conventional cultivars and their three isogenic transgenic derivatives, in two locations, Lavras and Patos de Minas in Minas Gerais, Brazil, in the 2012/13 crop season. Data on yield (kg&middotha-1), plant height (cm), and tolerance to lodging were collected. The cultivars had different responses. The cultivar BRS/MG46 Conquista had lower yields than its isogenic derivative BRS Valiosa RR, but this was not seen with the conventional cultivar M-SOY 6101, which produced more than its isogenic derivative M-SOY 7211 RR. Between the isogenic cultivars BRS/MG68 Vencedora and BRS Favorita RR, there were no significant differences. Nevertheless, the contrast between the mean values of the transgenic and conventional cultivars shows that the conventional cultivars had mean values greater than the transgenic cultivars.

Allelic Relationship between <i>Lr</i>9 and the Leaf Rust Resistance Gene in Kharchia Local Mutant of Wheat

To confirm allelic relationship between Lr9 and the leaf rust resistance gene in KLM4-3B, genetics of resistance was studied using crosses (WL711 + Lr9) × WL711 and (WL711 + LrKLM4-3B) × WL711. The F2 populations in cross (WL711 + Lr9) × WL711 and (WL711 + LrKLM4-3B) × WL711 segregated in ratio of 3:1 for disease reaction at seedling stage against pathotype 77-5 of leaf rust. This suggests that rust resistance in these stocks are under the control of single dominant genes. Further, to study allelic relationship betweenLr 9 and LrKLM4-3B, F2 population of the cross (WL711 + LrKLM4-3B) × (WL711 + Lr9) was studied. A segregation ratio of 15:1 implies that the two genes Lr9 and LrKLM4-3B are non-allelic genes.

Preliminary evaluation of resistance genes in rice against bacterial leaf blight in Guilan Province—Iran

The reactions of rice bacterial leaf blight races were identified in Guilan province—Iran on 12 near-isogenic lines and 14 pyramiding lines from International Network for Genetic Evaluation of rice (INGER) and 8 local and improved Iranian varieties were evaluated under natural photoperiod condition in the field. Inoculation was done at panicle initiation by clipping the sterilized scissors in the bacterial suspension to booting stage. Scoring of inoculated plants was made 21 days after inoculation. Infection levels of pyramiding lines containing two to five resistance genes, expect, IRBB53 and IRBB61 with respectively resistance gene combination, Xa5 + Xa13 and Xa4 + Xa5 + Xa7, were not so clear. Among near-isogenic lines IRBB1, IRBB2, IRBB4 and IRBB10 carrying resistance gene Xa1, Xa2, Xa4 and Xa10 were susceptible;IRBB8, IRBB11, IRBB3, IRBB5 and IRBB13 were moderately susceptible;(having resistance gene Xa8, Xa11, Xa3, Xa5 and Xa13) IRBB14, IRBB21 and IRBB7 with respectively resistance gene Xa14, Xa21 and Xa7 were moderately resistance to bacterial blight. Furthermore, most of the time gene combinations support the strategy of pyramiding appropriate resistance gene. Local varieties were more susceptible than improved varieties to leaf blight disease. Among local varieties, Tarom was the most susceptible. And also, there were no significant differences among improved varieties and all of them were moderately resistance.

利用PDS系统实现配管设计的模型化

介绍了三维PDS系统在配管设计工作中的应用。利用这个系统逐步改进配管设计工作模式,逐步实现配管设计的模型化。

PDS ISO抽图机理及ISOBatch的二次开发

本文介绍了PDS批量抽取ISO图的原理和流程,以及基于其原理开发的二次软件ISOBatch。