转Bt基因棉Bt毒蛋白的ELISA检测方法的研究(英文)

利用纯化的Ｂｔ毒素蛋白（６７ｋＤａ）作为标准蛋白和免疫抗原,建立了检测转基因抗虫棉植株内Ｂｔ毒蛋白含量的抗体夹心ＥＬＩＳＡ方法。检测极限１μｇ·Ｌ－ １,线性范围１－ １５μｇ·Ｌ－ １,板内误差Ｃ．Ｖ．＜ ５％ ,板间误差Ｃ．Ｖ．＜ ５％ ,平均样品回收率９４．９４％ 。经稀释度试验等质量控制试验,表明所建立的ＥＬＩＳＡ方法是可靠的,可用于转基因棉花植株中Ｂｔ毒素蛋白含量的测定。     

Genetic analysis and genotype × environment (G × E) for grey leaf spot disease resistance in elite African maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) germplasm

Maize grey leaf spot (GLS) disease remains an important foliar disease in sub-Saharan Africa accounting for more than 25% yield losses in maize. Information on inheritance of GLS resistance of germplasm adapted to African environments is required in new sources being identified. Therefore, hybrids generated from a 10 × 10 half-diallel mating of tropical advanced maize inbred lines were evaluated in six environments to determine combining ability, genotype × environment interaction (G × E) and the impact of GLS disease on grain yield. General combining ability effects were highly significant and accounted for 72 and 68% of the variation for GLS resistance and grain yield, respectively. Significant specific combining ability effects associated with reduced disease levels were observed in some hybrids when one parent was resistant, and these may be exploited in developing single cross maize hybrids. Regression analysis showed a 260–320 kg ha^?1 decrease in maize grain yield per each increase in GLS disease severity score, and significant associations (r = ?0.31 to ?0.60) were observed between grain yield and GLS severity scores. This showed the potential of GLS disease to reduce yield in susceptible varieties grown under favourable disease conditions, without control measures. Genotype and genotype × environment biplots and correlation analysis indicated that the significant G × E observed was not due to changes in hybrid ranking, implying absence of a significant crossover interaction. Therefore, predominance of additive gene effects imply that breeding progress for GLS disease resistance would be made through selection and this could be achieved at a few hot-spot sites, such as Baynesfield and Cedara locations in South Africa, and still deploy the resistant germplasm to other environments in which they are adapted.     

Genetic analysis and associated SRAP markers for flowering traits of chrysanthemum (<Emphasis Type="Italic">Chrysanthemum morifolium</Emphasis>)

The inheritance of two flowering traits of chrysanthemum, initial blooming time and the duration of flowering, was investigated using segregation within an F ₁ population derived from a cross between the autumn-flowering ‘Yuhualuoying’ and the summer-flowering ‘Aoyunhanxiao’ cultivars. The analysis, based on a single segregating generation and the major gene plus polygene mixed inheritance model, showed that the inheritance of both traits was compatible with the presence of two pairs of major genes displaying additivity–dominance–epistasis, with additivity predominating. As the heritability of both pairs of major genes was high (initial blooming time ~65%, duration of flowering ~72%), it should be possible to select for both traits in early breeding generations. A marker-trait association analysis based on sequence-related amplified polymorphism (SRAP) genotyping uncovered 10 (initial blooming time) and 12 (duration of flowering) markers significantly associated with phenotype, cumulatively explaining, respectively, 46 and 54% of the variation. Some potentially useful markers were identified.     

Molecular markers for improving control of soil-borne pathogen <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in sugar beet

Fusarium spp. cause severe damage in many agricultural crops, including sugar beet, with Fusarium oxysporum historically being considered as the most damaging of all species. Sugar beet needs to be protected from this class of soil-borne pathogens in order to ensure an optimal sugar yield in the field. Genetic control of the disease is crucial in managing these pathogens. Identification of single nucleotide polymorphism (SNP) markers linked to resistance can be a powerful tool for the introgression of valuable genes needed to develop Fusarium-resistant varieties. A candidate gene approach was carried out to identify SNP markers linked to putative Fusarium resistance sources in sugar beet. Five resistant analogue genes (RGAs) were screened by means of high resolution melting (HRM) analysis in a set of sugar beet lines, considered as resistant and susceptible to Fusarium oxysporum. HRM polymorphisms were observed in 80% of amplicons. Two HRM polymorphisms were significantly associated with Fusarium resistance (P < 0.05). The amplicons that showed association were sequenced and two SNPs were identified. The association was further validated on 96 susceptible and 96 resistant plants using competitive allele-specific PCR (KASPar) technology. The selected SNPs could be used for marker-assisted breeding of Fusarium resistance in sugar beet.     

Reaction of <Emphasis Type="Italic">Phaseolus</Emphasis> spp. genotypes to ashy stem blight caused by <Emphasis Type="Italic">Macrophomina phaseolina</Emphasis>

Ashy stem blight (ASB) caused by Macrophomina phaseolina (Tassi) Goidanich (Mp) is a devastating seed-transmitted disease in common bean in the tropics. The identification of resistant cultivars throughout the cropping season contributes to disease management. Resistance is found in the primary and tertiary gene pools. Our objectives were to determine (1) the reaction of Phaseolus spp. genotypes to two Mp isolates at vegetative and reproductive stages, (2) the area under disease progress curve (AUDPC), and (3) resistant plants per genotype at harvest. Twenty-three genotypes from different origins were planted in the greenhouse in 2016 and 2017. One less-aggressive Mp (PRJD16) and one more-aggressive (PRI16) isolate were inoculated one and three times, respectively, by the cut-stem method. ‘Beníquez’, ‘Othello’, and ‘Verano’ were highly susceptible (mean scores >8.0, and AUDPC values from 264.6 to 300.8) to both isolates. BAT 477 and NY6020-4 were intermediate (5.6 and 6.2; AUDPC: 161.6 and 187.1) to PRJD16 and susceptible (7.4 and 8.2; AUDPC: 209.4 and 235.1) to PRI16. Resistant genotypes (mean scores ≤3) were not identified in this study. However, A 195, ‘Badillo’, and ‘PC 50’ possessed lower mean scores (4.3–5.4) and AUDPC values (126.4–150.9) to both isolates. Furthermore, A 195 had the highest percentage of resistant plants (55.6%) followed by PC 50, I9365-31, and PI 321637 (27.8%) to PRJD16 at harvest. Thus, the identification of resistant parents across Phaseolus species is necessary to increase the levels of ASB resistance in common bean cultivars throughout the entire cropping season.     

Prediction of parental combination for introduction of stay-green associated loci in wheat

In wheat, the increase of yield and stability associated traits can be achieved by combining parents containing the stay-green trait and favorable alleles for grain yield. The aim of this work was to analyze the genetic dissimilarity between wheat lines from stay-green and synchronized maturation groups and elite cultivars. Moreover, to propose promising combinations seeking the selection of high-grain yield and high bread-making quality genotypes containing stay-green trait. The experiment was conducted in a randomized block design with three replications in 2003, 2004, and 2005, using sister-lines with the presence and absence of stay-green trait and elite cultivars. Genetic variability exists among wheat strains from the synchronized stay-green maturation group and elite cultivars. Genotypes of maturation group stay-green obtained an average performance superior to the synchronized group. Crosses between stay-green lines and the CEP 29 and BRS 177 cultivars are promising in the selection of genotypes carrying the stay-green trait with high yield and bread-making quality.     

Are tree breeders properly predicting genetic gain? A case study involving <Emphasis Type="Italic">Corymbia</Emphasis> species

The estimation of quantitative genetic parameters in breeding programs is important to ensure efficient selection. In this context, knowledge of the mating system is critical, as it underpins assumptions about inter-relatedness on which variance component estimation depends. However, proper account of the breeding system is not always taken, either because it is unknown and/or because it is ignored. That eucalypts have a mixed-mating system is well-established, however many breeders use models that assume outcrossed mating with an infinite number of male parents (i.e. allogamous mating), from which genetic parameter estimates are then used to predict genetic gains. First-generation, open-pollinated progeny tests of Corymbia citriodora subsp. citriodora and C. citriodora subsp. variegata, being managed for seed production, were used to investigate the likely bias, resulting in overestimation of genetic parameters under the assumption of allogamous mating. When we assumed allogamous mating, we observed inflated predictions of additive variance and narrow-sense heritability (\(\hat{h}_{a}^{2}\)) of diameter at breast height and height. The overestimate of \(\hat{h}_{a}^{2}\) was approximately 32% for C. citriodora subsp. citriodora and 21% for C. citriodora subsp. variegata. Inappropriate modelling of relatedness in eucalypts that assumes panmictic outcrossing when in fact these species have a mixed-mating system results in overestimates of the population genetic gain with selection.     

Comparative Studies on the Role of Organic Biostimulant in Resistant and Susceptible Cultivars of Rice Grown under Saline Stress - Organic Biostimulant Alleviate Saline Stress in Tolerant and Susceptible Cultivars of Rice

Plants are sessile organisms that experience various abiotic stresses during their lifespan and try to adapt to these environmental stresses by manipulating their physiological, biochemical, cellular, and molecular mechanisms. Salinity is one of the important abiotic stress that affects the metabolism and physiology of plant cells that leads to serious damage to crops and productivity. We investigated the response of two contrasting (salt susceptible and tolerant) cultivars during saline stress by modulating its effect with the application of an important natural biostimulant panchagavya (PG). The results showed that the salinity stress greatly influenced and negatively affects the plant growth, biochemical attributes, and induces the expression of various genes in both cultivars. Furthermore, we assessed the effect of PG alone and by amending with NaCl to alleviate the saline stress which showed a significant enhancement of biochemical and physiological characteristics in both cultivars. Furthermore, we assessed the response of seven autophagy associated gene (ATG1, ATG3, ATG4, ATG6, ATG7, ATG8, and ATG9), BAX Inhibitor -1 (BI-1), Mitogen activated Protein Kinase–1 (MAPK-1), WRKY53, Catalase -1 (CAT-1), Superoxide Dismutase (SOD), and Glutathione Peroxidase (GPX) genes in rice that displayed the differential expression pattern during saline stress in both cultivars. We concluded that saline stress can be manipulated by the application of PG and positively regulate the physiological, biochemical, and gene expression response in salt-susceptible and -tolerant rice cultivars. Furthermore, the current study also suggested that salinity is a mutifactorial and multigenic response. Autophagy and programmed cell death regulated along with salinity and was helpful in adapting the tolerance against the stress condition.     

Identification of quantitative trait locus and prediction of candidate genes for grain mineral concentration in maize across multiple environments

Trace metal elements are essential in daily diets for human health and normal growth. Maize is staple food for people in many countries. However, maize has low mineral concentration which makes it difficult to meet human requirements for micronutrients. The objective of this study was to identify quantitative trait locus (QTL) and predict candidate genes associated with mineral concentration in maize grain. Here, a recombinant inbred line population was used to test phenotype of zinc (Zn), iron (Fe), copper (Cu) and manganese (Mn) concentrations in six environments and then a QTL analysis was conducted using single environment analysis along with multiple environment trial (MET) analysis. These two strategies detected a total of 64 and 67 QTLs for target traits, respectively. Single environment analysis revealed 13 QTL bins distributed on seven chromosomes. We found that five candidate genes associated with mineral concentration were located in the same intervals identified by Comparative mapping of meta-QTLs in our previous study. The genetic and phenotypic correlation coefficients were depended on the nutrient traits and they were significant between Fe and Zn, Fe and Cu, Fe and Mn in all environments. The results of this study illustrated the genetic correlation between maize grain mineral concentrations, and identified some promising genomic regions and candidate genes for further studies on the biofortification of mineral concentration in maize grain.     

Evaluation of potato clones for their adaptation to medium altitude conditions in the tropics

Potato is one of the most important crops in the world, including in the tropics. Potato requires low temperatures or cool climate for optimum tuber yield, which in the tropics, can be obtained mostly in high altitude growing areas. The limitation of land availability for potato production in highland areas of some tropical countries has steered the recent development of potato production to medium altitude areas. The objective of our study is to identify potato accessions with promising levels of heat resistance in laboratory and with good adaptation to medium altitude in the field. We used in vitro assay to screen a number of potato clones with possible resistance to heat stress on the basis literature and database of the International Potato Center, supplemented with the collection of Bogor Agricultural University. We then continued with verification of selected clones using field test in high altitude (1300 m above sea level) and medium altitude (700 m above sea level) areas. Our result showed that there is considerable variation of S. tuberosum for their adaptation to medium altitude areas in the tropics that can be exploited in breeding programs. We identified one genotype PKHT 2013-06 that was superior for its ability to produce tubers under extreme conditions. Thus, it might be very suitable to be used in a breeding program to develop tolerant varieties of potato to medium altitude areas in the tropics.