Identification of CMS maintainers and restorers for hybrid rice development in Ghana

The cytoplasmic-genetic male sterility is the most widely used system for hybrid rice (Oryza sativa) seed production. It requires three lines: the two hybrid parents (A and R lines) and a third line (maintainer (B line)), which maintains the sterility of the female parent during seed increase. Studies were undertaken to identify maintainers and restorers for introduced cytoplasmic male sterile (CMS) lines using simple sequence repeat (SSR) markers and testcrosses. Molecular screening with SSR marker RM6100 grouped the assembled inbred lines into 30 potential restorers and 55 potential maintainers. One maintainer (WAS 127 -12-1-6-3-1) and 16 restorers were confirmed by testcrosses. WAS 127-12-1-2-3 had higher grain yield per plant in crosses with CMS1 (IR69625A) than the local checks (AgraRice and Jasmine 85) and could be used as a restorer to develop a locally adapted, high-yielding hybrid. Conversion of WAS 127 -12-1-6-3-1 into a local CMS line was stopped because of sterility instability in the BC₁F₁ generation. Continued testcrosses to the set of identified potential maintainers could help identify a stable one that can successfully be converted into a locally adapted CMS line.     

Ae显性突变体对玉米农艺性状及直链淀粉含量的影响分析

玉米直链淀粉是重要的食品和工业原料,高直链淀粉性状的产生受多种酶和基因-环境等因素的共同影响。玉米ae基因（amylose extender）突变导致子粒中直链淀粉含量增高、子粒表面皱缩、淀粉颗粒呈不规则形状、尺寸不均匀且表面基质蛋白增多,用于玉米育种后获得的品种直链淀粉含量升高,产量显著降低。通过将显性Ae突变体与16份优质玉米自交系杂交,对F₁代植株进行开花期、株型、产量性状调查,分析F₁代子粒总淀粉、直链淀粉的含量。结果表明,Ae突变体与不同的自交系杂交,F₁代直链淀粉含量及产量的变化存在明显差异,与基因型有密切关系,以Ae突变体和W16F41、CA240为亲本杂交,可显著提高直链淀粉含量同时产量变化最小。根据F₁代表现筛选直链淀粉含量提高、产量变化较小的双亲配制杂交种,可加速高直链淀粉玉米新品种培育进程。     

Selection before backcross during exotic germplasm introgression

Introgression of genes from exotic germplasm into breeding populations can broaden the genetic base of crop improvement. Only a very small percentage of genetic variability has been used in crop breeding programs. Traditionally, F₁ plants are used to backcross to the adapted lines or populations. An alternative approach is to backcross the F₂ individuals selected for agronomic acceptability. Our objective was to determine whether selection before backcross would lead to more progenies with both high yield and acceptable levels of agronomic performance than direct backcross without selection. To test the feasibility of the proposed approach, we conducted parallel experiments in which two exotic sorghum accessions were crossed to two adapted sorghum parents and further backcrossing was conducted with either F₁ or selected F₂ plants. Fifty random S₁ families were evaluated in three test environments. Although selection before backcross resulted in a higher frequency of families with maturity equal to or earlier than those of the adapted parents, no consistent changes in grain yield and plant height were observed between populations with and without selection. Similar results were found with either an inbred or a population as the recurrent parents. Given these findings and the extra generation required, we do not recommend selection before backcross in the process of introgression of exotic germplasm.     

不同种质基础玉米材料诱导效率及加倍效率研究

以不同类群骨干自交系PH4CV、Mo17、PH6WC、郑58、D299、H29、丹黄34、丹598、昌7-2和S121为基础试材,按照群内和群间杂交共组配15个基础群体,分别形成F₁、F₂、F₃和F₄共4个自交世代,研究其单倍体诱导率和加倍率。利用高频诱导系丹诱3号对骨干自交系和不同遗传基础的自交后代进行单倍体诱导,利用浓度为0.06%的秋水仙素对各世代单倍体子粒进行加倍处理。结果表明,不同种质类群自交系间诱导率存在显著差异,以Lancaster群代表系PH4CV、Mo17和Reid群代表系PH6WC、郑58诱导率较高。高诱导率类群通过群内和群间组配基础试材,同样具有较高的诱导率,同时利用杂种优势提升了LRC、SPT和PB群骨干系的诱导率。诱导率随着自交世代增加而显著降低,在F₂世代加倍效果最好。     

Increasing protein content and digestibility in sorghum grain with a synthetic biology approach

Despite great genetic diversity, sorghum grain consistently suffers from poor protein digestibility. The physicochemical packaging of protein bodies which consist of protease-resistant β- and γ-kafirin is considered a major obstacle. A synthetic β-kafirin gene, which shares the endosperm-specific promoter and signal peptide with the native β-kafirin gene (Sobic.009G001600.1), was transformed into sorghum inbred line Tx430. The gene was modified with ten additional proteolytic sites. These sites were designed to be amenable to cleavage by pepsin and/or chymotrypsin proteinases. Five independent transgenic lines were regenerated by microprojectile transformation. Notably, considerably more protein was observed in the peripheral endosperm of transgenic lines under scanning electron microscopy. Microscopy revealed invaginated or irregularly shaped protein bodies in the endosperm of transgenic lines. Grains of transgenic lines contained 11–37% more protein, which was 11–21% more pepsin digestible and 7–25% more chymotrypsin digestible than Tx430. Additionally, the abundant synthetic β-kafirin protein (5.6% of total protein) was detected by mass spectrometry data analysis in the transgenic line 9-1. Field-grown homozygous transgenics retained higher protein content, larger seed size and no reduction in grain number per plant. The results illustrated that plant synthetic biology could play an important role in improving sorghum nutritional value.     

Correlation studies on 1000-true-seed weight,tuber yield and other morphological traits in potato (Solanum tuberosum L.)

Summary Simple phenotypic, genotypic and environmental correlation coefficients were calculated from one year's data for nine characters in F₁ seedlings of 72 crosses and 18 parents of potato (Solanum tuberosum L.). Crosses were made in a line x tester model having 12 female lines and six male testers. The experiment was designed as a randomized block replicated three times at Kufri, Simla (India) during the 1979–80 crop season. Positive phenotypic and genotypic correlations of 1000-TPS weight were found with total tuber yield, number of leaves on the main shoot, number of shoots per plant and harvest index. The results suggest that 1000-TPS weight could be used when attempting to select for yield and other characters in F₁ seedlings. Adapted from a thesis submitted and approved for the degree of Doctor of Philosophy of Agra University.     

Rubisco small subunits of C4 plants,Napier grass and guinea grass confer C4-like catalytic properties on Rubisco in rice

ABSTRACT

Overexpression of Rubisco small subunit (RbcS) of C₄ plant, sorghum (sorghum bicolor) was shown to enhance the catalytic turnover rate (k _cat) of Rubisco in rice (Oryza sativa). In this study, the effects of other Rubisco small subunits of C₄ plants, Napier grass (Pennisetum purpureum) and guinea grass (Megathyrsus maximus) on kinetic properties of Rubisco in rice were studied. The expression levels of Napier grass RbcS (NgRbcS) and guinea grass RbcS (GgRbcS) proteins accounted for 41% and 45% of total RbcS, respectively in homozygous overexpression lines. The k _cat and K _m for CO₂ (Kc) of Rubisco were increased in all transgenic lines. Interestingly, the k _cat was markedly higher in NgRbcS homozygous line, whereas K _c was notably higher in GgRbcS homozygous line. Although its effects depend on species, these results suggest that the introduction of C₄ RbcS are effective approaches to alter the catalytic properties of Rubisco in rice.     

Towards F<Subscript>1</Subscript> Hybrid Seed Potato Breeding

Compared to other major food crops, progress in potato yield as the result of breeding efforts is very slow. Genetic gains cannot be fixed in potato due to obligatory out-breeding. Overcoming inbreeding depression using diploid self-compatible clones should enable to replace the current method of out-breeding and clonal propagation into an F₁ hybrid system with true seeds. This idea is not new, but has long been considered unrealistic. Severe inbreeding depression and self-incompatibility in diploid germplasm have hitherto blocked the development of inbred lines. Back-crossing with a homozygous progenitor with the Sli gene which inhibits gametophytic self-incompatibility gave self-compatible offspring from elite material from our diploid breeding programme. We demonstrate that homozygous fixation of donor alleles is possible, with simultaneous improvement of tuber shape and tuber size grading of the recipient inbred line. These results provide proof of principle for F₁ hybrid potato breeding. The technical and economic perspectives are unprecedented as these will enable the development of new products with combinations of useful traits for all stakeholders in the potato chain. In addition, the hybrid’s seeds are produced by crossings, rendering the production and voluminous transport of potato seed tubers redundant as it can be replaced by direct sowing or the use of healthy mini-tubers, raised in greenhouses.     

Genetic enhancement of Sorghum (Sorghum bicolor (L) Moench) for grain mould resistance: II. Breeding for grain mould resistance

Grain mould causes qualitative and quantitative loss to grain in sorghum. Grain mould resistance is a complex problem as grain mould is caused by complex of fungi and the resistance is governed by many traits. Breeding efforts during the last 3 decades to develop grain mould resistance in high yielding genotypes have not paid many dividends. We developed a strategy to breed for grain mould resistance in high yielding back ground. Twenty five crosses between elite lines and grain mould resistant genetic stocks (susceptible × resistant/moderately resistant and moderately resistant × resistant crosses) were evaluated in F₁, and derivatives performing superiorly for grain mould resistance in F₂-F₄ at physiological maturity were advanced. The early generation material F₂s (10) and F₃s (125) in 6 locations (representing rainy-season-sorghum growing 6 states of India where grain mould is one of the major biotic stresses), and later generations F₄s and F₅s in 3 locations (one location, Parbhani is a hot spot for grain moulds and 2 locations, Hyderabad and Coimbatore in epiphytotic conditions) were evaluated. Only 25 selections out of 384 derivatives in F₄ were superior over locations for grain mould resistance at physiological maturity and harvest maturity (Our simultaneous studies in RILs for grain mould resistance across years and locations have shown that the variation obtained for grain mould resistance at physiological maturity is genetically governed and the grain mould score further gets compounded at harvest maturity depending on rainfall received after physiological maturity). These superior lines were advanced and further evaluated in F₅ and F₆ for grain mould resistance and grain yield. During 2007, out of 25 F₅ derivatives, 12 were on par (scored 3.1-4.4) with resistant check, B 58586 (3.2 score) where as susceptible check, 296 B registered a score of 7.5. GMN nos. 41, 52, 59, and 63 performed on par with resistant check, B 58586 for grain mould resistance over 9 environments. Since we selected for grain mould resistance in early generations at physiological maturity in multi-locations, we could identify superior lines for grain mould resistance. Most of these lines are high yielding and on par with elite check, C43 for grain yield. These lines are distinct for DUS testing traits from grain mould resistant check, B 58586.     

水稻亚种间杂种优势利用中广亲和性的研究

 从1986年始对水稻广亲和性的资源筛选，亲和谱，遗传和利用进行了研究。试验结果表明：1）在籼粳杂种后代材料及美国稻品种中存在较丰富的广亲和品种，如T984，嘉40，Pecos和L201等均表现有良好的亲和性。2）各广亲和品种亲和谱存在明显的差异。T984，中国91，热研1号和嘉40的亲和谱较宽；而Dourada Percos和中作180的亲和谱较窄，即所谓的特写亲和性品种。3）广亲和品种的亲和性遗传亦有不同的表现。热研1号的广亲和性（以南京11和76-27bB作为籼粳测验种）受一对S_n⁵基因控制，其位于第6染色体山，与色素原基因C的交换值为4.71%；Pecos和L201的广亲和性（以南京11和秋光为测验种）除受色素原基因C连锁的主效基因控制外，还存在微效基因的修饰；T984的广亲和性（以辐8-1和秋光为测验种）与色素原基因C相互独立。此外，广亲和品种/粳型测验种的F2植株的亲和性强弱与相应的F3株系的平均结实率高低存在一定的关系。     

温带高油玉米自交系与热带玉米自交系配合力分析

采用Griffing Ⅵ 对6个国内温带高油系和4个热带系进行组配,将组配的45个杂交组合在云南省昆明、德宏两种不同的生态环境下进行农艺性状配合力分析和遗传参数估算。结果表明：热带系YML146、CML166、CML161、CML171是综合性状配合力表现较优良的种质材料,在高油玉米生产及育种上具有较大的利用潜力,是拓宽温带高油玉米种质遗传基础较好的种质资源;产量特殊配合力(SCA)效应值为正值的组合多数为温带高油系×热带系和热带系×温带高油系,说明温带高油玉米种质与热带种质具有较大的遗传差异,该组合模式具有较大利用价值。主要农艺性状的加性方差均大于非加性方差,说明这些性状主要表现加性遗传效应,可在早期世代进行选择。     

玉米禾谷镰孢穗腐病抗性的QTL定位

以3个高抗玉米自交系承351、丹598、吉V203和1个高感自交系ZW18为亲本,分别构建3个F₂群体及其对应的F_2:3家系,通过对F_2:3家系发病果穗进行图像处理的方法鉴定抗病表型,对禾谷镰孢穗腐病抗性进行QTL定位。3个F₂群体共检测到11个与禾谷镰孢穗腐病抗性有关的QTLs,分别可解释4.87%～40.98%的表型变异率。来源于抗病亲本承351的QTL-qRgr7-1位于7.02 bin,可解释高达13.76%～40.98%的表型变异率。通过与前期病害评级方法定位到的QTLs相比,qRgr7-1在Bins7.02上和qRger7.1完全重合,qRgr2-1在Bins2.01-2.02上和qRger2.1完全重合,qRgr10-1在Bins10.01-10.02上和qRger10.1完全重合。基于图像分析定位到的qRgr9-1、qRgr1-2、qRgr3-1分别与基于抗病评级定位到的区间存在重叠区域。利用不同方法定位到了相同的QTLs,一方面说明基于图像分析进行表型鉴定有一定的准确性,另一方面也验证了这些QTLs位点的真实性。     

Physio-morphological Studies of F1 Hybrids in Rice (Oryza sativa L.): Photosynthetic ability and yield

Summary

Photosynthetic ability in terms of CO₂ exchange rate and leaf area, dry matter accumulation (dry weight) and other important growth components along with yield and yield contributing characters in two F₁ hybrids of Akebono × Chiyonishiki (A × C) and Zenith × Akebono (Z × A) were studied. The photosynthetic rate at the flowering stage was lower than that at the panicle initiation stage in all the inbred parent cultivars and F₁ hybrids. Heterosis in photosynthetic rate was higher at the panicle initiation stage than that at the flowering stage. Pre-anthesis leaf area in the F₁ hybrid was significantly larger than that in the parent cultivars. However, the post- anthesis decline in leaf area was more rapid in the F₁ hybrids than in the parent cultivars and its magnitude was the highest in the F₁ hybrid of Z × A which was tall having long and curved panicles. Pre-anthesis dry matter accumulation in both hybrids was significantly higher in the F₁ plants than the parental cultivars, but post-anthesis dry matter accumulation in the F₁ hybrid was lower than the mid-parental value in Z × A. The degree of heterosis in grain yield varied with the hybrid combination, an average heterosis being 1.03. Heterosis in grain yield was closely associated with heterosis in harvest index.     

Quantitative genetics of rice IV. A breeding strategy

A systematic and objective breeding strategy is proposed, involving three stages of selection carried out at the F₁, F₃ and F₆ generations of pedigrees founded on single crosses between inbred parents. The first two of these stages involve selection between pedigrees and the third, selection within the pedigrees that survive the first two stages of scrutiny. The key features of this procedure are: that selection within pedigrees is delayed until individuals are effectively homozygous; that all characters are measured directly; and that decisions are based on the performance of plants that are raised in replicated and randomised trials. The replacement of the pedigree method by the new method should allow breeders to recognise and exploit the genetical variation between and within their crosses for characters of interest more efficiently than hitherto at an appreciably lower cost. While this new method has been proposed in response to a specific problem with rice, it is applicable to all crop plant species where the end products of breeding programmes are inbred lines.     

Flowering time in oat: Genotype characterization for photoperiod and vernalization response

Flowering time is an important component in the adaptation of oat to sub-tropical environments. Genotypes differ in their response to photoperiod and to vernalization. The objectives of this study were to identify photoperiod insensitive oat genotypes and then investigate the response of different oat genotypes to a period of vernalization (cold treatment), and to evaluate F₆ recombinant inbred lines from two oat crosses in two different environments (day-length increases and day-length decreases). The genotypes used in the studies were from the UFRGS Oat Breeding Program: UFRGS 8, UFRGS 881971 and UFRGS 930605 and from the University of Minnesota, USA: Amagalon/*Ogle, Coker 492/Starter-1 and PC68/5*Starter, and F₆ lines from the crosses UFRGS 8 × UFRGS 930605 and UFRGS 8 × PC68/5*Starter. UFRGS 8 showed no photoperiod sensitivity by flowering early regardless of the photoperiod length. Late flowering in the winter and early flowering in the summer characterize day-length-dependent genotypes like PC68/5*Starter. UFRGS 881971 responded to vernalization. Genotypes showed variability that can be used in the selection of lines/varieties that more effectively use the complete season.     

Analysis on genetic contribution of agronomic traits to total sugar in flue-cured tobacco (Nicotiana tabacum L.)

To uncover the genetic contributions of agronomic traits to content of total sugar (TS) and find indicator traits for indirect selection on TS in the flue-cured tobacco (Nicotiana tobacum L.), multivariable conditional analysis was conducted based on a genetic model containing additive–dominance and their interactions with environments. Fourteen cultivars (or breeding lines) and derived 41 F₁ crosses were grown at four locations in Yunnan province, China. Significant phenotypic contribution to TS was detected for six agronomic traits, plant height (PH), girth of stem (GS), internode length (INL), number of leaves (NL), length of middle leaves (LML) and width of middle leaves (WML). There was large contribution of additive effects due to each of the five agronomic traits (PH, GS, INL, LML and WML). The contribution ratio of dominance effect was high due to PH. By serving as high contributor of additive effects to TS and having high ratios of additive variance to phenotypic variance, INL and PH could be used as indicative agronomic traits for selecting breeding lines with suitable TS. Among the six agronomic traits, PH had the highest contribution to dominance effects of TS for most F₁ crosses, and could be used for selecting the crosses with suitable TS.     

小黑麦杂交F_1代真假杂种的ISSR标记鉴定及遗传多样性分析

为了给ISSR标记技术应用于小黑麦杂种鉴定提供参考,利用ISSR标记对小黑麦品系P1和P_2有性杂交得到的F_1代群体进行分子鉴定,同时结合田间表型调查结果对F_1代群体进行遗传多样性分析。结果表明,66个杂交F_1代单株中,52个为真杂种,其中,5个单株无主茎、未结实,1个生长后期死亡。14条ISSR引物对46个正常结实真杂种扩增的多态性条带百分率为61.19%,其中,UBC822的多态性条带百分率为100%,UBC808和UBC815的多态性条带百分率均为75%。F_1代群体和父母本的遗传相似系数为0.62~0.94,说明杂交后代产生了丰富的遗传变异。聚类分析将杂交F_1代群体和父母本在阈值0.748处分为3大类群,第1类群包括1份材料(父本,P1),第2类群包括1份材料(母本,P_2),第3类包括46个杂交F_1代单株;杂交F_1代与父母本的遗传距离较远,先与母本聚为一类,之后与父本聚为一类。田间调查结果表明,7个性状中,除株高偏向于父本外,分蘖数、有效分蘖数、穗长、小穗数、穗粒数和穗粒重均偏向于母本,与父本的差距较大,这与ISSR分析结果基本相吻合。     

Transgressive variants for red pericarp grain with high yield potential derived from Oryza rufipogon × Oryza sativa: Field evaluation, screening for blast disease, QTL validation and background marker analysis for agronomic traits

Five transgressive variants (advanced breeding lines from BC₂F₅ and BC₂F₆ generation) were derived from a cross between the wild relative, O. rufipogon Griff. and O. sativa L. subsp. indica cv. MR219, a popular high yielding Malaysian rice cultivar. The aim of the study was to evaluate the pericarp colour of the grains along with yield potential and to validate quantitative trait loci (QTLs) for agronomic traits. The variants were screened against blast disease. Background marker analysis was also done for the promising variants. The field trials were carried out at a single location (due to containment purposes) over two seasons using randomized complete block design (RCBD) with three replications. A trait-based marker analysis was used to identify QTLs for validation in BC₂F₅ generation. Analysis of variance (ANOVA) showed that the seasonal factors influenced different agronomic traits. Variant G33 produced significantly (p < 0.05) higher yield (5.20 t/ha) than the control, MR219 (4.53 t/ha). Eighteen QTLs for different agronomic traits were identified in BC₂F₂ population in a previous study. Among them 14 QTLs were found in BC₂F₅ population of the present study. The yield of variant G33 was influenced by several QTLs viz. qGPL-1, qSPL-1-2, qSPL-8 and qYLD-4, which were introgressed from the donor parent revealed by background marker analysis using BC₂F₇ generation. Percentage (99%) of red pericarp grain of G33 and G34 in BC₂F₅ and BC₂F₆ generations indicated the stability of pericarp colour which was transferred from the wild relative. Variant G33 showed resistance against two pathotype of blast disease (Magnaporthe oryzae). Among the evaluated variants, G33 could be considered for inclusion in the cultivar development program for red rice with high yield potential and resistance to blast disease. This study demonstrated that the alleles from wild relative could improve the yield and yield related traits through allelic interaction, even though the phenotypic traits were inferior to the recurrent parent.     

A New Dioic Acid from a wbl Gene Mutant of Deepsea-Derived Streptomyces somaliensis SCSIO ZH66

The wblA_so gene functions as a global regulatory gene in a negative manner in deepsea-derived Streptomyces somaliensis SCSIO ZH66. A new dioic acid (1) as well as two known butenolides (2 and 3) were isolated from the ΔwblA_so mutant strain of S. somaliensis SCSIO ZH66. The structure of 1 was elucidated by a combination of spectroscopic analyses, including MS and NMR techniques. In the cell growth inhibitory evaluation, compound 3 exhibited moderate activity against the human hepatic carcinoma cell line (Huh7.5) with an IC₅₀ value of 19.4 μg/mL, while compounds 1 and 2 showed null activity up to 100 μg/mL.     

Aspergillus Sydowii Marine Fungal Bloom in Australian Coastal Waters,Its Metabolites and Potential Impact on Symbiodinium Dinoflagellates

Dust has been widely recognised as an important source of nutrients in the marine environment and as a vector for transporting pathogenic microorganisms. Disturbingly, in the wake of a dust storm event along the eastern Australian coast line in 2009, the Continuous Plankton Recorder collected masses of fungal spores and mycelia (~150,000 spores/m³) forming a floating raft that covered a coastal area equivalent to 25 times the surface of England. Cultured A. sydowii strains exhibited varying metabolite profiles, but all produced sydonic acid, a chemotaxonomic marker for A. sydowii. The Australian marine fungal strains share major metabolites and display comparable metabolic diversity to Australian terrestrial strains and to strains pathogenic to Caribbean coral. Secondary colonisation of the rafts by other fungi, including strains of Cladosporium, Penicillium and other Aspergillus species with distinct secondary metabolite profiles, was also encountered. Our bioassays revealed that the dust-derived marine fungal extracts and known A. sydowii metabolites such as sydowic acid, sydowinol and sydowinin A adversely affect photophysiological performance (F_v/F_m) of the coral reef dinoflagellate endosymbiont Symbiodinium. Different Symbiodinium clades exhibited varying sensitivities, mimicking sensitivity to coral bleaching phenomena. The detection of such large amounts of A. sydowii following this dust storm event has potential implications for the health of coral environments such as the Great Barrier Reef.