Genetic Relation Analysis on Ramie[Boehmeria nivea(L.)Gaud.]Inbred Lines by SRAP Markers

The objective of this article is to reveal the variations of ramie inbred lines in DNA level and discuss their molecular background to provide a theoretical basis for ramie cross breeding. In the present study, the genetic relationships among 33 inbred line accessions and two wild types that originated from China and Brazil were estimated using sequence-related amplified polymorphism （SRAP） markers. The results showed that 33 out of 81 primer combinations turned out to be polymorphic and 332 polymorphism bands were obtained. On the basis of the appearance of the markers, the genetic relationships were analyzed using unweighted pair-group method of arithmetic average cluster analysis （UPGMA）, and the genetic Jaccard similarity coefficients were calculated. The inbred-lines originating from China and Brazil formed a cluster suggesting a possibility that the Brazilian cultivars could have developed from cultivars introduced from China. Within ramie inbred-lines, the groupings also indicated that the greatest genetic relationship among cultivars was correlated to the region of origin of cultivars. The results provided the evidence that SRAP was an efficient approach, suitable for taxonomic analysis of ramie inbred lines, To the authors＇ knowledge, this is the first application of SRAP marker on the systematics of ramie inbred lines.

Genetic analysis and QTL mapping of stalk cell wall components and digestibility in maize recombinant inbred lines from B73 × By804

The cell wall composition and structure of the maize stalk directly affects its digestibility and in turn its feed value.Previous studies of stem quality have focused mostly on common maize germplasm,and few studies have focused on high-oil cultivars with high grain and straw quality.Investigation of the genetic basis of cell wall composition and digestibility of maize stalk using high-oil maize is desirable for improving maize forage quality.In the present study,a high-oil inbred line(By804)was crossed as male parent with the maize inbred line B73 to construct a population of 188 recombinant inbred lines(RILs).The phenotypes of six cell-wall-related traits were recorded,and QTL analysis was performed with a genetic map constructed with SNP markers.All traits were significantly correlated with one another and showed high broad-sense heritability.Of 20 QTLs mapped,the QTL associated with each trait explained 10.0%–41.1%of phenotypic variation.Approximately half of the QTL each explained over 10%of the phenotypic variation.These results provide a theoretical basis for improving maize forage quality by marker-assisted selection.

Genetic analysis and QTL mapping of growth period traits and plant height traits in soybean recombinant inbred lines from Dongnong 47 × PI 317334-B

High and stable yield is the main goal of soybean genetic improvement.In this study,association analysis was used to detect the quantitative trait loci(QTL)for the plant height,and soybean growth period using 182 SSR markers in the RIL population of 136 F_(4:8) lines,which developed from a cross between photoperiod-insensitive cultivar‘Dongnong 47’and photoperiod-sensitive variety PI317334–B.The results showed that 33 QTLs related to soybean growth period and plant height traits were detected by compound interval mapping,and were located on 12 linkage groups including N,C1,C2,J,D1a,B2,E,G,A2,O,L,I,with the contribution rate of 7.85–33.84%.These QTL loci and linkage markers related to soybean photoperiod sensitivity,would be helpful to identify key genes that control soybean photoperiod sensitivity,and provide an important basis for the breeding of new photoperiod-insensitive soybean varieties based on molecular design breeding.

Genetic Diversity for Yield and Yield Contributing Characters of Short Statured Maize Inbred Lines

Genetic diversity is one of the useful tools to select appropriate lines for hybridization. Twenty short-statured maize inbred lines were taken for present study, which were collected from CIMMYT India and Mexico through Plant Breeding Division, Bangladesh Agricultural Research Institute (BARI), Gazipur. This experiment was conducted from November 2015 to April 2016 to identify parental lines to produce single cross short statured maize hybrids. From the genetic diversity studies, the importance of both additive and non-additive gene actions for the expression of yield and yield contributing characters were found. Values of vector I and II revealed that both the vectors had positive values for date of silking, plant height, rows/cob, grains/row and yield. These results indicated that these five characters had highest contribution towards divergence. Based on the relative magnitude of D2 values;20 inbred lines were grouped into five clusters. Seven inbred lines were selected on the basis of genetic diversity and per se performance which will be crossed separately in a half-diallel fashion to develop hybrids.

Determination of the Number of SSR Alleles Necessary for the Analysis of Genetic Relationships Between Maize Inbred Lines

The amount of molecular marker information has considerable impact on the results of studies of crop germplasm genetic relationships in crop. The number of alleles required to reveal genetic relationship in maize inbred lines is a theoretical issue that needs to be addressed. In this study, 112 pairs of SSR （simple sequence repeat） primers and 97 maize inbred lines were selected to study the relationship between the number of inbred lines and the number of SSR primers and alleles required for a stable cluster. The results showed that the number of SSR primers is not tightly associated with the stability of the cluster analysis results, while an increase in the number of alleles can significantly improve the stability of cluster analysis results. The number of inbred lines （X） is significantly associated with the number of alleles required for stable cluster analysis （Y）, and the regression equation is Y- 600.8xe（-15.9/x）. This equation can be used to calculate the number of SSR alleles required for a genetic relationship study of maize inbred lines. These results provide a reference for determining of SSR alleles number in genetic relationship analysis of maize inbred line and other crop germplasm.

Factors influencing seed reserve utilization during seedling establishment in maize inbred lines

Strong seedlings are essential for high yield.To explore the foundation of strong seedlings,we investigated various factors influencing the conversion and distribution of seed storage reserves during seedling establishment in maize inbred lines.Three maize inbred lines were used to explore the effects of seed size,seed vigor,illumination duration,temperature,water content,and salt concentration of the seedling medium on the utilization of seed storage reserves during seedling establishment.The results showed that the conversion rate of small seeds was 3.69 to 17.71%higher than that of large seeds.Moreover,prolonged illumination time was conducive to the formation of strong seedlings.However,low temperature,drought stress and salt stress reduced the conversion rate of seed storage reserves and increased the root/shoot ratio.These results could be used to guide field management during seedling emergence and develop improved germplasm with a high conversion rate of seed storage reserves.

Genetic Diversity Analysis of 13 Maize Inbred Lines by SSR Molecular Markers

This paper used SSR molecular markers to perform the genetic diversity analysis on 13 ordinary inbred lines of maize of different sources in order to divide heterotic groups of maize inbred line and predict heterosis. Using 12 pairs of SSR primers,a total of 47 allelic variants were detected in 13 inbred lines,2-5 alleles were detected for each pair of primers,an average of 3. 9,and polymorphism information content varied from 0. 379 to 0. 828. According to the cluster analysis,the 13 inbred lines could be divided into 5 groups.

Breeding Technology of Sunflower Inbred Lines with Four Generations in One Year

The breeding technology of sunflower inbred lines with four generations in one year was explored through the research on the biological characteristics of sunflower itself and the breeding status of sunflower inbred lines using the young embryo culture technology,the greenhouse seedling technology and field test planting,based on sunflower field breeding and planting combined with greenhouse planting.The reasonable selection of planting sites as the core of the breeding technology of sunflower inbred lines with four generations in one year is a guarantee.Embryos can be taken 7-10 d after pollination to enter the next generation of cultivation,thus shortening the generational cultivation cycle.The establishment of a breeding method of sunflower inbred lines with four generations in one year reduces the cost of breeding and has a practical role and significance for the field of sunflower inbred line breeding technologies.

Quantitative trait locus analysis of lateral branch-related traits in cucumber(Cucumis sativus L.)using recombinant inbred lines

A group of 224 recombinant inbred lines (RILs) was derived from a narrow cross between 2 cucumber (Cucumis sativus L.) lines, namely, S94 (Northern China type with weak lateral branch growth potential and early lateral branch sprouting time) and S06 (Northern European type with strong lateral branch growth potential and late lateral branch sprouting time). These lines were then used for investigating lateral branch-related traits. A total of 36 quantitative trait loci (QTLs) were detected for the following 4 lateral branch-related traits: lateral branch average length (LBAL), lateral branch total length (LBTL), lateral branch number (LBN), and first lateral branch node (FLBN). Further, each QTL explained 3.1% (lbtl2.1, spring) to 32.3% (lbn2.3, spring) of the observed phenotypic variance. Eleven QTLs (lbal1.1, lbtl1.1, lbn1.2, flbn1.2, etc.) for different traits were found to be clustered on the e23m18d-ME23EM6c section (7.4 cM) of linkage group (LG) 1; further, 15 QTLs (lbal2.1, lbtl2.1, lbn2.1, flbn2.1, etc.) were found to be clustered on the S94A1-ME4SA4a section (13.9 cM) of LG2. Twenty-one QTLs explained more than 10% of the phenotypic variance. Moreover, lbtl1.3 (autumn, 26.2%, logarithm of odds (LOD) = 17.4; spring, 26.9%, LOD = 17.9) had stable position and contribution in both seasons. Several se-quence-anchor markers (CMBR40, F, CS30, S94A1, CSWTA11B, etc.) were closely linked with some QTLs for LBAL, LBTL, LBN, and FLBN, which can be used for the marker-assisted selection to improve the plant architecture in cucumber breeding.

Evaluation of drought tolerance in ZmVPP1-overexpressing transgenic inbred maize lines and their hybrids

The vacuolar proton-pumping pyrophosphatase gene(VPP)is often used to enhance plant drought tolerance via genetic engineering.In this study,the drought tolerance of four transgenic inbred maize lines overexpressing ZmVPP1(PH4CV-T,PH6WC-T,Chang7-2-T,and Zheng58-T)and their transgenic hybrids was evaluated at various stages.Under normal and drought conditions,the height and fresh weight were greater for the four transgenic inbred maize lines than for the wild-type(WT)controls at the germination and seedling stages.Additionally,the transgenic plants exhibited enhanced photosynthetic efficiency at the seedling stage.In irrigated and non-irrigated fields,the four transgenic lines grew normally,but with increased ear weight and yield compared with the WT plants.Moreover,the ear weight and yield of the transgenic hybrids resulting from the PH4CV-T×PH6WC-W and Chang7-2-T×Zheng58-W crosses increased in the non-irrigated field.Our results demonstrated that the growth and drought tolerance of four transgenic inbred maize lines with improved photosynthesis were enhanced by the overexpression of ZmVPP1.Moreover,the Chang7-2 and PH4CV transgenic lines may be useful for future genetic improvements of maize hybrids to increase drought tolerance.

QTL mapping of grain appearance quality traits and grain weight using a recombinant inbred population in rice(Oryza sativa L.)

supported by a grant from the National High-Tech R＆D Program of China （2014AA10A603, 2014AA10A604）;a grant from the Youth Foundation in Sichuan, China （2011JTD0022）;the special fund for China Agricultural Research System （CARS-01-08）;the Provincial Specialized Funds for Innovation Ability Promotion in Sichuan, China （2013GXJS005）

Diversity, Structure, and Marker-Trait Association Analysis of the Maize Recombinant Inbred Line Population

Association mapping has emerged as a new tool to elucidate complex quantitative trait loci in maize, but there are few reports about systematic association analysis for the specific SSR markers with agronomic traits of interest in China. We investigated the morphological and genetic diversity and population structure for 76 maize recombinant inbred lines, and then association analysis were further performed between 48 simple sequence repeat loci and 17 morphological traits, consisting of nine ear-related traits and eight other traits. The 48 SSR markers were screened out and further classified into two groups including a group of loci in regions harboring reported quantitative trait loci that affect ear shape and a group of markers distributing on the whole genome randomly. The result indicated that the population of recombinant inbred lines was structured, showing five subpopulations. Our association results revealed that there were 82, 59, and 40 significant associations detected by K-test, logistic regression, and both analysis, respectively. When the 17 traits were considered separately, the significant associations between Q-SSRs and E-traits were raised to 27.8%, whereas the other groups of combinations ranged between 2.3 and 6.3%. As the proportion of significant associations is higher among the Q-SSR subset of markers and the subset of traits related to ear shape than those for all of the other combinations, we conclude that this approach is valid for establishing true positive marker-trait relationships. Our results also demonstrated that association mapping could complement and enhance previous QTL information for marker-assisted selection.

Genetic Diversity of Chinese Temperate and Exotic Tropical, Subtropical Quality Protein Maize Inbreds by SSR Markers

Information on genetic relationship is of great value to maize (Zea mays L.) breeding. Theobjectives of this study were: 1) to classify 22 quality protein maize (QPM) inbreds intodifferent groups by using simple sequence repeats (SSR) markers, which included exotictropical, subtropical and domestic temperate QPM and normal maize inbreds; 2) to examine theconsistency of grouping results obtained from SSR, specific combining ability (SCA) analysis,and genetic backgrounds of these inbreds. A set of 39 polymorphic SSR primers was selected from70 primer pairs, which detected 136 alleles among the 22 lines. The mean polymorphisminformation content was 0.55. Based on analysis of genetic similarities, five groups wereidentified including Luda Red Cob, Sipingtou, Reid, Lancaster and a miscellaneous group withseveral tropical inbreds which could not be classified into the above four groups. The resultsgenerally agreed with previous results based on analysis of yield combining ability andpedigree data.

Identification of the miniature pig inbred line by skin allograft

Skin grafting has been used as one of the most reliable tests to determine the genetic stability of laboratory animal such as mice and rats inbred line, but no identification of swine inbred lines by skin grafting has been reported. At present, Wuzhishan miniature pig （WZSP） inbred line has acquired the F24 individuals in China. In order to verify whether WZSP inbred line had D^en cultivated successfully, allogeneic skin grafts and related research were performed on F20 individuals of WZSP inbreeding population, compared with a control group of autologous transplantation. We observed the transplant recipients＇ wounds, detected peripheral blood-related indicators interleukin-2, 4 and 10, CD4~ and CD8~ lymphocytes, and conducted hematoxylin-eosin （HE） and Masson＇s staining of skin to judge whether the immune rejection reactions occurred within 28 days after transplantation. Chr. 7 genomic heterozygosity of 48 WZSP individuals from F20 to F22 was analyzed by high-density single nucleotide polymorphism （SNP） chips （60 000 SNPs）. The result showed that there were no significant differences in graft skin, the plasma interleukin-2, 4, 10, CD4~ and CD8~, HE and Masson＇s staining results between the allograft and autograft groups, and no immune rejection occurred on the allograft group. We found that 11 genes in Chr. 7 of major histocompatibility complex （MHC） I and MHC II were homozygous which confirmed that immune antibody of the allograft and autograft groups were highly identical and also provided a theoretical basis to no immune rejection occurred on the allograft in the inbred WZSP. The result proved that the WZSP inbred line had been cultivated successfully for the first time in the world. The test methods also provide a scientific basis for the identification of swine and mammal inbred lines.

Transfer and Detection of barstar Gene to Maize Inbred Line 18-599 (White) by Particle Bombardment

In China, the purity of maize hybrid strain is discomforting to the development of seed industrialization. Finding a new method for reproduction of maize hybrid strain is necessary. In this study, using particle bombardment, barstar gene was transferred into maize inbred line 18-599 （White）, which is an antiviral and high quality maize inbred line. By molecular detection of the anther of transgenic maize, two plants transferred with barstar gene were gained in this study, which are two restorer lines. The two plants showed normal male spike, and lively microspores. But the capacity of the two restorer lines should be studied in the future. The aim of this study is to find a new method of reproduction of maize hybrid strain using engineering restorer lines and engineering sterility lines by gene engineering technology.

Low-Temperature Response to Major Agronomic Traits by Using Recombinant Inbred Line(RIL) Populations Derived from Towada × Kunmingxiaobaigu

Development of the recombinant inbred line populations （RILs） is important basis to detect QTLs for cold tolerance at booting stage in rice. A set of 230 RILs derived from the cross of Towada and Kunmingxiaobaigu were used for evaluation of low-temperature response on major agronomic traits of plant height （PH）, panicle length （PL）, panicle exsertion （PE）, spikelet fertility （SF）, specific spikelet fertility （SSF）, and spikelets per panicle （SPP） under natural low-temperature growing environments in Yunnan Province, China. The results showed PH, PE, and SPP were mainly attributed by genotypes. PL was mainly influenced interactively by the genotypes × environments. SF and SSF were mainly controlled by the environments. Under the five different growth environments, F values of the six agronomic traits mentioned above ranged from 4.019 to 97.284. Significant difference was revealed between the lines. Under every environment, it indicated significantly positive correlation between SF and SSF, with correlation coefficients ranged from 0.826 to 0.885. It indicated significantly positive correlation between PH, PL, and PE. Under five different growing environments, variation coefficients of the six characters ordered in SSF （66.3%） 〉 PE （57.4%） 〉 SP （37.2%） 〉 SPP （16.2%） 〉 PH （9.6%） 〉 PL （6.4%）. SSF, PE and SF were most sensitive to low temperature stress at booting stage, while SPP, PH and PL being least. The RILs of Towada/ Kunmingxiaobaigu can be used as a genetic population to investigate cold tolerance at booting stage. SSF, PE and SF are most sensitive to cold tolerance at booting stage in rice. So far the the variation of PH, PL, and SPP related to cold tolerance are not clear under natural low-temperature environment. More tested environments and years are required to identify and evaluate cold tolerance at booting stage in rice.

A Dynamic Model of Heavy Ion ~7Li Irradiation Mutagenesis Based on Maize Inbred Line Nutrition Difference

To reveal the saddle-type dose effect relationship, we propose a radiation mutagenesis model based on maize nutrition difference resulting from heavy ion ~7Li radiation. Through irradiation mutagenesis, apparent trait selection, amino acids and fatty acids content determination, and modeling, dynamic evolution from microscopic damage and repair initiation to the final macroscopic biological effects are considered simultaneously. The results show that the steady state nature is independent of evolution time and only relates to different radiation doses.Heavy ion ~7Li radiation could effectively cause maize phenotypic variation and could improve nutritional quality.This model not only gives a good fit to the experimental results on most types of amino acids and fatty acids, but also offers an adequate explanation of the experimental phenomenon underlying the saddle-type bimodal dose effect. By combining experimental results with theoretical analyses, we suggest that the synergy of the stimulus effect and momentum transfer is the main cause of the saddle-type dose effect bimodal curve. This provides an effective strategy for conducting maize germplasm innovation.

Identifying Genomic Regions Conferring Morphological and Yield-related Traits in Soybean Based On A Four-way Recombinant Inbred Line Population

Improvement of seed yield of soybean(Glycine max(L.)Merr.)is generally achieved by combining morphological and yield-related traits,such as plant height(PH),node number on main stem(NN),pod number per plant(NP),seed number per plant(NS),100-seed weight(HSW)and seed weight per plant(SWPP).Identifying quantitative trait loci(QTLs)for morphological and yield-related traits is therefore important for breeding.In this study,a four-way recombinant inbred line population comprising 160 lines derived from the cross(Kenfeng14×Kenfeng15)×(Heinong48×Kenfeng19)was planted in five different environments and morphological and yield-related trait data were used to identify QTLs by the inclusive composite interval mapping method.Totally 38 QTLs for PH,40 QTLs for NN,26 QTLs for NP,10 QTLs for NS,26 QTLs for HSW and 49 QTLs for SWPP were detected in 125 genomic regions.Single QTLs explained 2.17%-14.60%,2.00%-10.04%,2.37%-9.77%,2.62%-8.61%,0.47%-6.51%and 0.14%-12.39%of the phenotypic variation for PH,NN,NP,NS,HSW and SWPP,respectively.Among these 125 genomic regions,120 were newly associated with morphological and yield-related traits.The results would facilitate the molecular breeding of morphological and yield-related traits in soybean.

Dissection of two QTL clusters underlying yield-related heterosis in the cabbage founder parent 01–20

Cabbage has significant heterosis and most commercial cultivars are hybrids.To explore genetic basis of cabbage heterosis and promote cabbage heterosis utilization,we constructed two populations by crossing 100 DH lines derived from a cabbage hybrid 01–20×96–100 with two female parents.Hybrids exhibited different extents of heterosis,the mean value of economic yield was 2.6 times bigger than parents.We identified 66 and 73 QTLs associated with mid-parent heterosis and transgressive heterosis of twelve yield-related traits,respectively.Some QTLs could be detected under the two-year experiment existed in two populations with different testers,showing relatively high phenotypic contribution rate(15.8%–20.0%).Heterosis QTLs exhibited clustered distribution in several cabbage chromosome regions.Two dominant genetic regions,mk300–316 and mk258–268,originated from the elite parent 01–20,exhibited significant genetic effects for yield-related heterosis,which were first identified.Three elite DH lines(D22,D46,D83)harboring these two dominant regions were selected as having strong heterosis in cabbage production.Candidate gene analysis revealed that some genes participating in biosynthetic processes of carbohydrates and some responses to auxin might affect cabbage yield heterosis.QTL identification and genetic dissection of yield-related traits provide new insights into the genetic effects of cabbage heterosis.

水稻耐淹性状QTL分析

利用籼稻品种贵9B和粳稻品种热研2号作为亲本,构建100个贵9B/热研2号重组自交系(recombinant inbred lines, RILs)家系为作图群体,进行水稻耐淹QTL检测及其遗传效应分析。以胚芽鞘长度为水稻耐淹强弱的表型数据,运用完备区间作图定位法,在第6和第9条染色体上检测到2个水稻耐淹QTL,依次命名为qGS-6-1、qGS-9-1,2个位点的LOD值分别为2.89、2.51,贡献率分别为12.03%、10.52%,qGS-6-1等位基因源于贵9B,qGS-9-1等位基因源于热研2号。该研究可为克隆控制耐淹的基因以及培育适宜直播的水稻品种提供重要的理论依据。