Validation of PyMBMS as a High-throughput Screen for Lignin Abundance in Lignocellulosic Biomass of Grasses

Pyrolysis molecular-beam mass spectrometry (PyMBMS) was tested as a high-throughput method for relative abundances of guaiacyl and syringyl lignin in lignocellulosic cell-wall materials from stems of a population of maize intermated B73?×?Mo17 (IBM) recombinant inbred lines. Variations of up to twofold across the population in phenylpropanoid abundance were observed. Several histochemical and quantitative biochemical assays were used to validate the mass spectrometric data for lignin, hydroxycinnamic acids, crystalline cellulose, non-cellulosic glucans, and xylans. We demonstrate PyMBMS to be a valid high-throughput screen suitable for analysis of lignin abundance in large populations of bioenergy grasses. Pentose from xylans and hexose from cellulosic and non-cellulosic glucans also varied substantially across the population, but abundances of diagnostic fragments for these monosaccharides were not well correlated with the abundance of cell-wall polysaccharides.     

Genetic Determinants of the Network of Primary Metabolism and Their Relationships to Plant Performance in a Maize Recombinant Inbred Line Population

Deciphering the influence of genetics on primary metabolism in plants will provide insights useful for genetic improvement and enhance our fundamental understanding of plant growth and development. Although maize (Zea mays) is a major crop for food and feed worldwide, the genetic architecture of its primary metabolism is largely unknown. Here, we use high-density linkage mapping to dissect large-scale metabolic traits measured in three different tissues (leaf at seedling stage, leaf at reproductive stage, and kernel at 15 d after pollination [DAP]) of a maize recombinant inbred line population. We identify 297 quantitative trait loci (QTLs) with moderate (86.2% of the mapped QTL, R² = 2.4 to 15%) to major effects (13.8% of the mapped QTL, R² >15%) for 79 primary metabolites across three tissues. Pairwise epistatic interactions between these identified loci are detected for more than 25.9% metabolites explaining 6.6% of the phenotypic variance on average (ranging between 1.7 and 16.6%), which implies that epistasis may play an important role for some metabolites. Key candidate genes are highlighted and mapped to carbohydrate metabolism, the tricarboxylic acid cycle, and several important amino acid biosynthetic and catabolic pathways, with two of them being further validated using candidate gene association and expression profiling analysis. Our results reveal a metabolite-metabolite-agronomic trait network that, together with the genetic determinants of maize primary metabolism identified herein, promotes efficient utilization of metabolites in maize improvement.     

High-throughput Screening of Recalcitrance Variations in Lignocellulosic Biomass: Total Lignin,Lignin Monomers,and Enzymatic Sugar Release

The conversion of lignocellulosic biomass to fuels, chemicals, and other commodities has been explored as one possible pathway toward reductions in the use of non-renewable energy sources. In order to identify which plants, out of a diverse pool, have the desired chemical traits for downstream applications, attributes, such as cellulose and lignin content, or monomeric sugar release following an enzymatic saccharification, must be compared. The experimental and data analysis protocols of the standard methods of analysis can be time-consuming, thereby limiting the number of samples that can be measured. High-throughput (HTP) methods alleviate the shortcomings of the standard methods, and permit the rapid screening of available samples to isolate those possessing the desired traits. This study illustrates the HTP sugar release and pyrolysis-molecular beam mass spectrometry pipelines employed at the National Renewable Energy Lab. These pipelines have enabled the efficient assessment of thousands of plants while decreasing experimental time and costs through reductions in labor and consumables.     

Exploring the Genetic Characteristics of Two Recombinant Inbred Line Populations via High-Density SNP Markers in Maize

Understanding genetic characteristics can reveal the genetic diversity in maize and be used to explore evolutionary mechanisms and gene cloning. A high-density linkage map was constructed to determine recombination rates (RRs), segregation distortion regions (SDRs), and recombinant blocks (RBs) in two recombinant inbred line populations (RILs) (B73/By804 and Zong3/87-1) generated by the single seed descent method. Population B73/By804 containing 174 lines were genotyped with 198 simple sequence repeats (SSRs) markers while population Zong3/87-1 comprised of 175 lines, were genotyped with 210 SSR markers along with 1536 single nucleotide polymorphism (SNP) markers for each population, spanning 1526.7 cM and 1996.2 cM in the B73/By804 and Zong3/87-1 populations, respectively. The total variance of the RR in the whole genome was nearly 100 fold, and the maximum average was 10.43–11.50 cM/Mb while the minimum was 0.08–0.10 cM/Mb in the two populations. The average number of RB was 44 and 37 in the Zong3/87-1 and B73/By804 populations, respectively, whereas 28 SDRs were observed in both populations. We investigated 11 traits in Zong3/87-1 and 10 traits in B73/By804. Quantitative trait locus (QTLs) mapping of SNP+SSR with SNP and SSR marker sets were compared to showed the impact of different density markers on QTL mapping and resolution. The confidence interval of QTL Pa19 (FatB gene controlling palmitic acid content) was reduced from 3.5 Mb to 1.72 Mb, and the QTL Oil6 (DGAT1-2 gene controlling oil concentration) was significantly reduced from 10.8 Mb to 1.62 Mb. Thus, the use of high-density markers considerably improved QTL mapping resolution. The genetic information resulting from this study will support forthcoming efforts to understand recombination events, SDRs, and variations among different germplasm. Furthermore, this study will facilitate gene cloning and understanding of the fundamental sources of total variation and RR in maize, which is the most widely cultivated cereal crop.     

Genetic Regulation of Mup Production in Recombinant Inbred Mice

Inbred strains of mice excrete all three major urinary proteins (mups) when induced by testosterone, but differ as to the relative proportions and total levels of each mup present. We have now determined the urinary mup phenotypes before and after testosterone treatment of seven recombinant inbred strains derived from progenitor strains exhibiting different mup phenotypes. The results confirm previous observations indicating that total control of mup protein production is a multigenic process. One locus, Mup-a on chromosome 4, determines the relative mup protein proportions after induction by testosterone. Mup-a, together with other genetic sites, determines the basal mup proportions. Genes other than Mup-a determine the kinetics of mup induction and total mup excretion.     

Identification of QTLs for Seed Dormancy in Cultivated Peanut Using a Recombinant Inbred Line Mapping Population

Plant Molecular Biology Reporter - Seed dormancy is an important breeding trait for the development of certain types of peanut cultivars. Peanut cultivars with seed dormancy can inhibit preharvest...     

Mapping Quantitative Trait Loci for Grain Appearance Traits of Rice Using a Recombinant Inbred Line Population

A linkage map consisting of 221 markers was constructed based on a recombinant inbred line (RIL) population from the cross between Zhenshan 97 and Minghui 63. Quantitative trait loci (QTL) mapping was carried out for grain appearance traits such as grain length, grain width and grain shape in rice in 1998 and 1999. Based on interval mapping method at the threshold LOD≥2.4, six, two and two QTLs were detected for grain length, grain width and grain shape, respectively, in 1998; In 1999, three, two and two QTLs were identified for the three traits, respectively. Of them, seven QTLs were simultaneously identified in both of the years. The QTL with large effects located in the interval RG393-C1087 on chromosome 3 not only controlled the grain length, but also influenced the grain shape. It explained 57.5%, 61.4% and 26.7%, 29.9% of phenotypic variation of the grain length and the grain shape in two years, respectively. The QTL with large effects located in the interval RG360-C734B on chromosome 5 affected the grain width and the grain shape. It explained 44.2%, 53.2% and 32.1%, 36.0% of phenotypic variation of the grain width and the shape in two years, respectively. Eight, five and five QTLs were identified for the grain length, width and shape, respectively, based on mixed linear-model composite interval mapping method at P =0.005. Their general contributions were 58.81%, 44.75%, and 57.47%. One QTL for the grain length was found to be significant interaction with environment.     

Inhibition of Maize Caffeate 3-O-Methyltransferase by Nitecapone as a Possible Approach to Reduce Lignocellulosic Biomass Recalcitrance

Plant Molecular Biology Reporter - Maize (Zea mays) caffeate 3-O-methyltransferase (ZmaysCOMT, EC 2.1.1.68), a key enzyme of the phenylpropanoid pathway, catalyzes the O-methylation of caffeic acid...     

利用水稻重组自交系群体定位谷粒外观性状的数量性状基因

用区间作图和混合线性模型的复合区间作图两种方法,对水稻（Oryza sativa L）珍汕97和明恢63组合的重组自交系群体的谷粒外观性状－粒长,粒宽和粒形进行了数量性状基因（QTL）定位,用区间作图法在LOD≥2．4水平上（近拟于a=0.005),1998年对粒长,粒宽和粒形分别检测到6,2放2个QTLs,1999年对以上3个性状分别检测到3,2和2个QTLs,其中7个QTLs在两年均检测到,位于第3染色体RG393－C1087区间的QTL效应大,同时影响粒长和粒形,两年贡献分别为57．5％,61．4％和26．7％,29．9％,位于第5染色体RG360－C734B区间的QTL效应大,同时影响粒宽和粒形,两年贡献率分别为44．2％,53．2％和32．1％和36．0％,用混合线性模型的复合区间作图法在P＝0．005水平上,对粒长,粒宽和粒形分别检测到8,5和5个QTLs,共解释各自性状变异的58．81％,44．75％和57．47％,只检测到1个QTL与环境之间存在的显互作。     

Detection of Quantitative Trait Loci for Yield and Drought Tolerance Traits in Soybean Using a Recombinant Inbred Line Population

To investigate the genetic basis of drought tolerance in soybean ( Glycine max L. Merr.) a recombinant inbred population with 184 F_2:7:11 lines developed from a cross between Kefeng1 (drought tolerant) and Nannong1138-2 (drought sensitive) were tested under water-stressed and well-watered conditions in field and greenhouse trials. Traits measured included leaf wilting coefficient, excised leaf water loss and relative water content as indicators of plant water status and seed yield. A total of 40 quantitative trait loci (QTLs) were identified: 17 for leaf water status traits under drought stress and 23 for seed yield under well-watered and drought-stressed conditions in both field and greenhouse trials. Two seed yield QTLs were detected under both well-watered and drought-stressed conditions in the field on molecular linkage group H and D1b, while two seed yield QTLs on molecular linkage group C2 were found under greenhouse conditions. Several QTLs for traits associated with plant water status were identified in both field and greenhouse trials, including two leaf wilting coefficient QTLs on molecular linkage group A2 and one excised leaf water loss QTL on molecular linkage group H. Phenotypic correlations of traits suggested several QTLs had pleiotropic or location-linked associations. These results will help to elucidate the genetic basis of drought tolerance in soybean, and could be incorporated into a marker-assisted selection breeding program to develop high-yielding soybean cultivars with improved tolerance to drought stress.     

Detection of Quantitative Trait Loci for Yield and Drought Tolerance Traits in Soybean Using a Recombinant Inbred Line Population

To investigate the genetic basis of drought tolerance in soybean （Glycine max L. Merr.） a recombinant inbred population with 184 F2：7：11 lines developed from a cross between Kefengl （drought tolerant） and Nannong1138-2 （drought sensitive） were tested under water-stressed and well-watered conditions in field and greenhouse trials. Traits measured included leaf wilting coefficient, excised leaf water loss and relative water content as indicators of plant water status and seed yield. A total of 40 quantitative trait loci （QTLs） were identified： 17 for leaf water status traits under drought stress and 23 for seed yield under well-watered and drought-stressed conditions in both field and greenhouse trials. Two seed yield QTLs were detected under both well-watered and drought-stressed conditions in the field on molecular linkage group H and Dlb, while two seed yield QTLs on molecular linkage group C2 were found under greenhouse conditions. Several QTLs for traits associated with plant water status were identified in both field and greenhouse trials, including two leaf wilting coefficient QTLs on molecular linkage group A2 and one excised leaf water loss QTL on molecular linkage group H. Phenotypic correlations of traits suggested several QTLs had pleiotropic or location-linked associations. These results will help to elucidate the genetic basis of drought tolerance in soybean, and could be incorporated into a marker-assisted selection breeding program to develop high-yielding soybean cultivars with improved tolerance to drought stress.     

ISSR分子标记技术在分析玉米自交系遗传关系研究中的适用性

利用ISSR分子标记技术对59份玉米自交系和1份大刍草材料进行遗传多样性分析.21对ISSR引物共扩增出475条不同位置的带,平均22.6条;多态性带469条,平均22.3条,百分率高达98.7%;不同引物的多态性信息指数(PIC)在0.84～0.94之间.60份材料的遗传相似系数在0.23～0.48之间.经聚类分析,可将这些材料分成两大组,共9个亚组,并且在一定程度上与血缘和系谱是一致的,也存在一些例外.结果表明,ISSR标记尽管可以用于进行遗传多样性分析,但并不是最好的分子标记类型.综合考虑不同的分子标记类型的优缺点,认为ISSR标记在遗传研究较少的作物上应用潜力较大.     

利用水稻重组自交系群体对一些重要农艺性状进行基因定位和互作分析

对水稻（OryzasativaL．）籼（indica）、粳（Japonica）交（窄叶青8号×京系17）通过一粒传获得了一个自交9代的重组自交系（RIL）群体,该群体含有107个稳定纯合的株系,通过构建分子连锁图谱,对水稻播抽期、株高、每穗颖花数、每穗实粒数、200粒重和结实率进行数量性状基因座位（QTL）分析,定位了影响播抽期的2个QTLs、株高的2个QTLs、每穗颖花数的2个QTLs和每穗实粒数的1个QTL。其中,控制播抽期的1个QTL即qHD－8为主效基因,控制株高的1个QTL即qPH－1为主效基因,其余为微效基因。同时分析了影响水稻生产力的数量性状基因座位间的互作。结果表明,影响播抽期、株高、每穗颖花数、每穗实粒数和结实率的互适型互作共有24个,对性状有6．2％～10．9％的贡献率。     

Rates of Spontaneous Mutation in Bacteriophage T4 Are Independent of Host Fidelity Determinants

Bacteriophage T4 encodes most of the genes whose products are required for its DNA metabolism, and host (Escherichia coli) genes can only infrequently complement mutationally inactivated T4 genes. We screened the following host mutator mutations for effects on spontaneous mutation rates in T4: mutT (destruction of aberrant dGTPs), polA, polB and polC (DNA polymerases), dnaQ (exonucleolytic proofreading), mutH, mutS, mutL and uvrD (methyl-directed DNA mismatch repair), mutM and mutY (excision repair of oxygen-damaged DNA), mutA (function unknown), and topB and osmZ (affecting DNA topology). None increased T4 spontaneous mutation rates within a resolving power of about twofold (nor did optA, which is not a mutator but overexpresses a host dGTPase). Previous screens in T4 have revealed strong mutator mutations only in the gene encoding the viral DNA polymerase and proofreading 3'-exonuclease, plus weak mutators in several polymerase accessory proteins or determinants of dNTP pool sizes. T4 maintains a spontaneous mutation rate per base pair about 30-fold greater than that of its host. Thus, the joint high fidelity of insertion by T4 DNA polymerase and proofreading by its associated 3'-exonuclease appear to determine the T4 spontaneous mutation rate, whereas the host requires numerous additional systems to achieve high replication fidelity.     

水稻叶片性状和根系活力的QTL定位

应用由247个株系组成的珍汕97B/密阳46重组自交系(RIL)群体及其分子标记连锁图谱,检测控制剑叶、倒二叶、倒三叶的5个形态性状和控制根系伤流量性状的数量性状座位(QTL)。在9个标记区间检测到控制叶片形态性状的24个QTL,LOD值为2．9～11．8,单个QTL的表型变异贡献率为4．0％～32．5％;分别检测到56对和4对控制叶片形态和根系活力的上位性互作,绝大多数互作发生在2个不表现加性效应的座位之间。与该群体产量性状QTL的研究结果相比较,发现控制叶片性状和根系活力的QTL与产量性状QTL往往处于相似的染色体区间。     

玉米耐盐种质筛选及群体遗传结构分析

本研究采用盆栽法评价了157份玉米自交系的苗期耐盐性,并利用115对SSR标记解析了耐盐自交系的群体遗传结构。结果表明,T5V、N1026、农大1145及4S等10份玉米自交系为高耐盐玉米种质;处理后10d株高、地上鲜重、地下鲜重、地上干重、地下干重及存活率可作为玉米苗期耐盐鉴定的重要指标;利用SSR分子标记,结合系谱资料,将157份自交系划为6个类群,其中具有通系5血缘（Ⅰ类群）、泰国糯玉米种质血缘（Ⅲ类群）及旅大红骨、黄早四等血缘（Ⅵ类群）的自交系耐盐性较强,是开展玉米耐盐育种的重要种质类群。本研究筛选到的耐盐种质将为玉米耐盐遗传机制研究、玉米种质遗传改良及耐盐分子育种提供优异的基因资源。     

贵州70份玉米自交系的SSR标记遗传多样性及其杂种优势群分析

从251个SSR标记中筛选出均匀分布在玉米基因组上的88个SSR标记,用以分析评价贵州省2000年以来47个审定品种的70份亲本材料的遗传多样性。SSR标记检测的结果:88个标记共检测出466个等位基因,每个标记可检测等位基因2～18个,平均为5.31个;每个标记位点的多态性信息量(PIC)变化为0.213～0.965,平均为0.586,这表明贵州玉米自交系具有较为丰富的遗传多样性。POPTREE聚类分析结果:70份自交系分为Ⅰ、Ⅱ和Ⅲ类群。Ⅰ类群含8个自交系,以瑞德和兰卡斯特等温带种质为主。Ⅱ类群有11个自交系,以PN78599、瑞德和兰卡斯特等温带种质为主。Ⅲ类群拥有51个自交系,可分为A和B 2个亚群,B亚群还可再分为B1和B2 2个次亚群,A亚群中的10个系以我国地方温带种质为主,B1次亚群中的19个系以贵州地方亚热带种质为主,B2次亚群中的22个系以泰国苏湾热带种质为主。杂种优势利用分析的结果表明,贵州近些年在玉米育种中,主要是利用贵州地方亚热带种质和泰国苏湾热带种质2个杂种优势群,这与其多态位点百分率较高有关,与其群内SSR位点的平均等位数较多有关。贵州玉米育种利用的种质类型较少,有必要加强玉米种...     

微卫星标记在玉米群体遗传多样性研究中的应用

简要综述了近几年利用微卫星(SSR)分子标记技术分析玉米群体遗传多样性过程中的研究进展对研究中混合取样,荧光标记技术,共享数据库等新方法的使用作了阐述.     

The Nxsm Recombinant Inbred Strains of Mice: Genetic Profile for 58 Loci Including the Mtv Proviral Loci

We report the construction of 17 recombinant inbred (RI) strains of mice derived from the progenitor strains NZB/BINRe and SM/J and the typing of this RI strain set, designated NXSM, for 58 loci distributed on 16 autosomes and the X chromosome. Two backcrosses involving NZB/BINJ and SM/J were constructed to confirm chromosomal assignments and determine gene orders suggested from NXSM RI strain data. From these results we recommend that chromosomal assignments and gene orders suggested from analyses of RI strain sets be confirmed using data obtained by other means. We also typed NZB/BINJ and SM/J for mammary tumor proviral (Mtv) loci. Both strains share three previously described Mtv loci: Mtv-7, Mtv-14 and Mtv-17. In addition, NZB/BINJ contains the previously described Mtv-3 and Mtv-9 loci and two new Mtv proviral loci: Mtv-27 located on chromosome (Chr) 1 and Mtv-28 located on the X chromosome. SM/J contains the previously described loci Mtv-6 and Mtv-8. Four LTR, mink cell focus-forming murine leukemia viral loci were identified and mapped: Ltrm-1 on Chr 12, Ltrm-2 on Chr 16, Ltrm-3 on Chr 5, and Ltrm-4 on Chr 13. The Tgn locus was positioned proximal to the Ly-6 locus on Chr 15.