Development of two multiplex PCR assays targeting improvement of bread-making and noodle qualities in common wheat

Wheat quality properties are genetically determined by the compositions of high and low molecular weight glutenin subunits, grain hardness, polyphenol oxidase (PPO) activity and starch viscosity. Two multiplex PCR assays were developed and validated using 70 cultivars and advanced lines from Chinese autumn‐sown wheat regions. Multiplex PCR I includes molecular markers for genes/loci ω‐secalin, Glu‐B1‐2a (By8), Glu‐D1‐1d (Dx5), Glu‐A3d, Glu‐B3 (for non‐1B·1R type) and Pinb‐D1b targeting improved gluten parameters and pan bread quality. Multiplex PCR II comprises markers for genes/loci Ppo‐A1, Ppo‐D1 and Wx‐B1b targeting improved noodle quality. The results were consistent with those achieved by SDS‐PAGE and RP‐HPLC, indicating that the two multiplex assays were highly effective, with good repeatability and low costs enabling their use in wheat breeding programmes. In total, nine alleles (subunits) at locus Glu‐B1, four at Glu‐D1 and five at Glu‐A3 locus were identified, and the alleles (subunits) Glu‐B1b (7 + 8), Glu‐B1c (7 + 9), Glu‐D1a (2 + 12), Glu‐D1d (5 + 10), Glu‐A3a, Glu‐A3c and Glu‐A3d were most frequently present in the cultivars and lines tested. The 1B·1R translocation was present in 28 (40.0%) lines, whereas the Wx‐B1 null allele for better noodle quality was present in only seven (10.0%) cultivars and advanced lines, and 37 (52.9%) lines had Pinb‐D1b associated with hard grains. The allele Ppo‐A1b on chromosome 2AL associated with lower PPO activity was present in 38 (54.3%) genotypes, whereas the less effective allele Ppo‐D1a on chromosome 2DL, also associated with low PPO activity was present in 45 (64.3%) of genotypes. These two multiplex PCR assays should be effective in marker assisted selection targeting improved pan bread‐making and noodle qualities.     

The effects of low doses of Senna occidentalis seeds on broiler chickens

The effects of 0.5%, 0.3% and 0.1% w/w concentrations of Senna occidentalis (So) seed mixed with commercial ration were studied in 18 groups of 32 broiler chicks each, from 1 day to 49 days of age. Three groups were fed one of the rations throughout their lives (TL). Three other groups were fed one of the rations from the 1st to the 28th day of life (starter phase, SP), and the final 3 groups were fed one of the rations from the 29th to 49th day (finisher phase, FP). Each experimental group was matched by a control group fed the same diet over the same period but without the inclusion of So. All the animals were killed at 49 days of age, and blood was collected from 10 birds in each group for biochemical studies (ALT, AST, GGT, LDH, UA). A complete necropsy was performed on 3 birds from each group. No significant differences in the biochemical parameters in the serum were found between the control and experimental chicks, but animals treated with 0.5% So in groups FP and TL, gained less weight and chicks that received 0.3% So or 0.5% So in the ration throughout life (TL) had a larger feed conversion ratio. Besides this, degenerative changes were found in the striated skeletal muscle in the chest, in the myocardium and in the liver in the animals that received the higher concentrations of So seeds.     

Study of the soil-plant (carrot)-animal cycle of nutritive and hazardous minerals in a rabbit model.

Carrots were grown on soils polluted by heavy metal salts. Each particular microelement reached a high concentration [molybdenum (Mo) 39.00, cadmium (Cd) 2.30, lead (Pb) 4.01, mercury (Hg) 30.00, and selenium (Se) 36.20 mg/kg dry matter] in the carrot. In a metabolic balance trial conducted with 15 male and 15 female New Zealand White rabbits, the control animals (n = 5) were fed ad libitum with concentrate as basal diet, while the other rabbits received the basal diet and carrots containing the particular microelement. Blood samples were taken to determine the activity of serum enzymes. To investigate the metabolism of Mo, Cd, Pb, Hg and Se, samples were taken from the heart, liver, lungs, kidneys, spleen, ovaries/testicles, entire digestive tract, adipose tissue, femur, hair, faeces and urine. Carrot had significantly higher digestibility for all nutrients than the rabbit concentrate. Carrot samples of high Pb content had the lowest digestibility of crude protein. The microelements differed in their rate of accumulation in the organs examined: Mo and Cd accumulated in the kidneys, Pb in the kidneys, liver, bones and lungs, Hg in the kidneys and liver, while Se in the liver, kidneys and heart. The proportions of microelements eliminated from the body either via the faeces and urine (Mo 80.18% and Se 47.41%) or via the faeces (Cd 37.86%, Pb 66.39%, Hg 64.65%) were determined. Pathohistological examination revealed that the rate of spermatogenesis was reduced in the Mo, Cd, Pb and Hg groups compared to the control. Lead, Cd and Hg intake resulted in a considerable decrease in gamma-glutamyltransferase (GGT) and in an increase of alkaline phosphatase (ALP) activity because of damages to the kidneys and bones. All experimental treatments decreased the activity of cholinesterase (CHE) because of lesions in the liver.     

小麦丛矮病研究之三:灰飞虱同发病的关系

1976—1977年河北晋县贺家寨大队小麦春季灰飞虱的传带病毒率为31—52%,不同地块同代灰飞虱的带毒率异差较大:而秋苗期则为12—35.4%,其中不同地块上同代(四代成虫)灰飞虱的带毒率的差异从33.3%至35.4%则不太大。灰飞虱的有效传毒指数(即虫口数×自然带毒率)同田间小麦发病率的相关性是非常显著的,其r=0.9769。1975—1976两年调查及计算分析结果,其直线回归公式如y=4.12x-2.577。在有效传毒指数0.7—20.9之间用于予测发病率有效。人工分期侵染试验证明小麦在生长发育过程中受侵愈早,病情愈重,其中死株率也愈大。秋季侵染的死株率达95.6%,病指为99.6;夏季侵染的死株率为0,病指为57.6。秋季小麦出苗前后的大量虫口形成小麦10月中下旬的一个发病高峰,第二年春季4月中旬的虫口高峰形成5月上旬的一个发病高峰。3月下旬的一个发病高峰是越冬前受侵的。病毒在小麦植株中的潜育期的长短随小麦的生长速度而异。温度不过其中因素之一,最短5—8天,最长30—40天。侵染后如小麦进入越冬期,即不表现症状,一直要到返青以后。介体灰飞虱在河北晋县以第五代3—4龄若虫在冬麦地及草荒中越冬。夏季在玉米上可以产卵并孵化,但不能长期存活,主要在地头沟边及夏作物荫蔽下的禾草上越夏。冬麦治虫防病的重点应在秋季麦苗出土前,春季则应在3月中旬左右。冬麦适当推迟播种有利于减少发病率。棉间作麦比平作的病情重六倍而粮麦间作的病情比平作的重九倍。     

A methodology to detect and quantify five pathogens causing potato tuber decay using real-time quantitative polymerase chain reaction

ABSTRACT Late blight (Phytophthora infestans), pink rot (Phytophthora erythroseptica), leak (Pythium ultimum), dry rot (Fusarium sambucinum), and soft rot (Erwinia carotovora subsp. carotovora and subsp. atroseptica) are particularly damaging diseases of stored potato tubers worldwide. In this study, we present a methodology to detect and quantify the causal agents of the five aforementioned diseases from whole potato tubers, using real-time quantitative-polymerase chain reaction. Six primer pairs were designed to amplify targets smaller than 150-bp DNA in single copy protein-coding gene targets of each of the pathogens and the potato host. Using a large collection of pure culture DNA samples, all primer pairs specifically detected the DNA target in the intended pathogenic species. Amplification efficiencies over a five-log dilution series ranged between 95 and 100% and were unaffected by the presence of large amounts of host DNA. The detection level of the primers reached 0.5 pg of target DNA. Pathogens were detected in 100 pg of total DNA extracted from 170 to 250 g of tubers, 4 days after inoculation, regardless of the presence of symptoms. The presence of P. erythroseptica, Pythium ultimum, or E. carotovora was also detected in 1 ng of DNA extracted from potato tubers collected from a commercial storage facility. This study provides the first step in a methodology to predict the storability of potato tubers following harvest.     

Improved Cold and Drought Tolerance of Doubled Haploid Maize Plants Selected for Resistance to Prooxidant tert‐Butyl Hydroperoxide

To improve the abiotic stress tolerance of maize (Zea mays L.), doubled haploid (DH) plants were produced by in vitro selection of microspores exposed to tert‐butyl hydroperoxide (t‐BuOOH) as a powerful prooxidant This study investigated the tolerance of the progenies of t‐BuOOH‐selected DH lines to oxidative stress, cold and drought in controlled environment pot experiments by analyses of photosynthetic electron transport and CO₂ assimilation processes, chlorophyll bleaching and lipid peroxidation of leaves. Our results demonstrated that the t‐BuOOH‐selected DH plants exhibited enhanced tolerance not only to oxidative stress‐induced by t‐BuOOH but also to cold and drought stresses. In addition, they showed elevated activities of antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, catalase, glutathione reductase and glutathione S‐transferase when compared with the DH lines derived from microspores that were not exposed to t‐BuOOH and to the original hybrid plants. The results suggest that the simultaneous up‐regulation of several antioxidant enzymes may contribute to the oxidative and cold stress tolerance of the t‐BuOOH‐selected DH lines, and that the in vitro microspore selection represents a potential way to improve abiotic stress tolerance in maize.     

Ultrastructure of a new type of sensory ending in Echinostoma revolutum cercaria (Trematoda: Echinostomatidae)

Two pairs of hitherto unknown multiciliate sensory endings have been demonstrated in free-swimming cercariae of Echinostoma revolutum using transmission and scanning electron microscopy. This type of receptor consists of a bundle of 36--40 short, conically projecting cilia ensheathed by a collar formed by a circular evagination of tegument in shape of a cup. The cilia projecting from the centre of a widened nerve fibre, filled with electron-lucid vesicles, possess a well developed basal body and do not contain a ciliary rootlet. The function of this receptor is discussed.     

Crossability and cytological studies in Solanum macrocarpon and Solanum linnaeanum (Solanaceae).

Summary Crossing experiments involving the various groups of Solanum macrocarpon L. complex, including wild (dasyphyllum), semi-wild and cultivars produced fully fertile F1 and F2 hybrids. This confirmed earlier findings that these groups belong to the same biological species. S. macrocarpon complex and S. linnaeanum Hepper ex Jaeger are isolated by reproductive barriers. F1 and F2 hybrids between various groups of S. macrocarpon showed heterosis. F2 superior hybrids were isolated as candidates for the future breeding programmes. Most wild-type traits like prickliness, hairiness were dominant to those possessed by cultivars. Relatively resistant hybrids to pest attack were noted. S. macrocarpon complex and S. linnaeanum are diploids (2n=24). The overall nature of the karyotype suggests that the two are related. The F1 hybrids between the cultivars of S. macrocarpon and the wild group (S. dasyphyllum) showed normal meiosis. A lack of isolating barriers confirmed that S. macrocarpon complex constitute a single biological species.