A Simple Method for the Mating of Sclerotinia trifoliorum

Rehnstrom, A. L., and Free, S. J. 1993. A simple method for the mating of Sclerotinia trifoliorum. Experimental Mycology 17, 236-239. A simple method which allows for the controlled mating of L and S mating type strains of Sclerotinia trifoliorum is described. Using the method, we have been able to mate L and S strains and have demonstrated the segregation of the genetic markers involved in mycelial incompatibility into the progeny.     

Construction of a bio-test for infecting red clover plants with Sclerotinia trifoliorum

Sclerotinia trifoliorum causes clover cancer in red clover crops. Clover cancer is difficult to control and completely resistant red clover varieties are not available. Breeding for resistant red clover varieties is being slowed down because little is known about the diversity of European S. trifoliorum populations and because of the lack of bio-tests that are useable in breeding programs. The first objective of this research was to develop a reliable high-throughput bio-test, useable in breeding programs. The second objective was to optimise another bio-test, based on isolated leaves, for more precise studies. First, we optimised a method for ascospore production of S. trifoliorum. Once produced, the ascospores were used to evaluate the effects of climate conditions, ascospore concentration and plant age on the high-throughput bio-test. For the bio-test on isolated leaves, the effects of infection method, incubation conditions, incubation period, ascospore concentration, leaf growth stage and mechanical damage were evaluated. In the high-throughput bio-test, disease levels rose with increasing ascospore concentration up to 20,000 spores/ml. The plant age had a small, yet significant effect on the disease level. For the isolated leaf bio-test, the most effective and most repeatable infection method was spraying of an ascospore suspension. Disease levels continued to increase with rising concentrations and incubation time did not interact with plant susceptibility levels. The youngest completely opened leaf yielded the most repeatable results. Both bio-tests were shown to be correlated and could be valuable instruments for breeding programs and for studying plant-pathogen interactions.     

A Comparison of Laboratory Tests in the Malabsorption Syndrome

Absorption of radioiodinated triolein was tested in 93 individuals (15 controls, 28 cases of functional diarrhea, and 50 patients with suspected or manifest malabsorption syndrome).The results so obtained were compared in 59 cases with estimations of fecal radioactivity and chemically determined fecalfat excretion. A highly significant correlation was found.The urinary xylose excretion test was performed in 96 individuals. The diagnostic value of the radioactive fat absorption test was compared with that of the xylose excretion test in 78 cases. Both tests were diagnostic in cases of non-tropical sprue, but the xylose excretion test reflected the clinical improvement more accurately in treated cases. The radioactive fat absorption test was more reliable in the diagnosis of malabsorption secondary to bowel resection than was the xylose excretion test, but the converse was true in the diagnosis of malabsorption secondary to gastrectomy. The simultaneous use of these two tests was found to be a simple and reliable screening procedure.     

Strategies for Improving Drought Resistance in Grain Legumes

This review distills recent information on drought resistance characteristics of grain legumes with a view toward developing appropriate genetic enhancement strategies for water-limited environments. First, the possible adaptations that allow grain legumes to better cope with drought stress are summarized. It is suggested that there are considerable gains to be made in increasing yield and yield stability in environments characterized by terminal drought stress by further exploiting drought escape, by shortening crop duration. Many traits conferring dehydration avoidance and dehydration tolerance are available, but integrated traits, expressing at a higher level of organization, are suggested to be more useful in crop improvement programs. Possible genetic improvement strategies are outlined, ranging from empirical selection for yield in droughted environments to a physiological genetic approach. It is suggested that in view of recent advances in understanding drought resistance mechanisms, the latter strategy is becoming more feasible. It is concluded that use of this recently derived knowledge in a systematic manner can lead to significant gains in yield and yield stability of the world's major grain legumes, as they are mainly grown (and will continue to be grown) under rain-fed conditions.     

Agrobacterium rhizogenes Mediated Transformation of the Forage Legumes Medicago sativa and Onobrychis viciifolia

Three cultivars of M. sativa and one cultivar of O. viciifoliawere evaluated for their response to inoculation with A. rhizogenesstrain A4T (containing pRiA4b). A cultivar-dependent responsewas observed in M. sativa with 94%, 25%, and 4% of infectedstem explants producing transformed roots in the cultivars Vertus,Regen-S, and Rangelander, respectively. In O. viciifolia cv.Hampshire Giant, an explant-dependent response was observedwith 78% and 50% of seedling cotyledon and hypocotyl explantsresponding, respectively. Leaf explants failed to produce transformedroots. Transformed roots showed plagiotropic and negativelygeotropic growth on hormone-free agar MS medium. Productionof transgenic shoots from O. viciifolia root cultures occurredspontaneously. Recovery of transgenic plants from M. salivacv. Rangelander was achieved by transfer of callus (inducedon UM medium containing 2·0mg dm^–3 2,4-D and 0·25mg dm^–3 kinetin) to MS medium containing 0·5 ingdm^–3 BAP and 0·05 mg dm^–3 NAA. Cultured rootsof both species synthesized opines (agropine and mannopine).Extensive morphological variation was observed in plants ofM. sativa (clone Al) and O. viciifolia (clone A4Tl) establishedin the glasshouse. DNA sequences homologous to TL-DNA and TR-DNAwere present in root clones and regenerated plants. Key words: Agrobacterium rhizogenes, Medicago sativa, Onobrychis viciifolia, transformed roots, transgenic plants     

Simple Method for the Separation of Ascospores

A simple method for the separation of ascospores is described. To isolate single spores from adhesive ascospores and the mycelium, the suspension was sucked through a combination of sintered-glass plates with different pore sizes.     

Somatic Hybrids of the Forage Legumes Medicago sativa L. and M. falcata L.

Protoplasts isolated from embryogenic cell suspensions of tetraploidMedicago sativa cv. Europe (2n= 4? = 32) and M. falcata (2n=4? = 32) were fused using polyethylene glycol (PEG). Heterokaryonswere isolated by micromanipulation and cultured in the presenceof nurse protoplasts from albino or embryogenic cell suspensionsof M. sativa, to give free-floating embryos and embryogeniccalli. Ninety-nine plants were regenerated from somatic embryos.Fifteen of the plants exhibited leaf abnormalities and did notsurvive transfer from culture to the glasshouse. The remainingphenotypically normal plants were established ex vitro and floweredat maturity. Morphological and biochemical analyses confirmedthat 12 of the phenotypically normal plants were somatic hybrids.Morphological characteristics of the hybrids, including plantstature, internode length, leaf size, flower colour, and podshape, were intermediate compared with those of the purple floweredM. sativa and yellow flowered M. falcata parents. Flowers ofthe hybrids were yellow traced with purple-blue veins. Isoenzymebanding patterns for esterase showed bands additional to thoseof M. sativa and M. falcata. The chromosome complements of individualhybrids varied from (2n= 4? = 32) to 58.     

天然氨基酸物质的不同制造技术的比较研究

依据 1 7～ 1 8种天然氨基酸物质的三种制造技术的比较研究 ,无论是基于生物资源还是基于其产品销售和环境保护方面 ,都证明了蛋白质水解制造法优于发酵法和合成法     

Diversity study on Sclerotinia trifoliorum Erikks., the causal agent of clover rot in red clover crops (Trifolium pratense L.)

Since the 16th century, red clover has been an important crop in Europe. Since the 1940s, the European areal of red clover has been severely reduced, due to the availability of chemical fertilizers and the growing interest in maize. Nowadays there is a growing interest in red clover again, although some setbacks still remain. An important setback is the low persistence of red clover crops. Clover rot, caused by the ascomycete fungus Sclerotinia trifoliorum Erikss., is a major disease in Europe and reduces the persistence of red clover crops severely. The fungus infects clover plants through ascospores in the autumn, the disease develops during the winter and early spring and can kill many plants in this period. In early spring, black sclerotia, serving as surviving bodies, are formed on infected plants. Sclerotia can survive up to 7 years in the soil (Ohberg, 2006). The development of clover rot is highly dependent on the weather conditions: a humid fall, necessary for the germination of the ascospores and an overall warm winter with short periods of frost are favourable for the disease. Cold and dry winters slow the mycelial growth down too much and prevent the disease from spreading. Clover rot is difficult to control and completely resistant red clover varieties have yet to be developed. Because of the great annual variation in disease severity, plant breeders cannot use natural infection as an effective means to screen for resistant material. Breeding for resistant cultivars is being slowed down by the lack of a bio-test usable in breeding programs. When applying artificial infections, it is necessary to have an idea of the diversity of the pathogen. A diverse population will require resistance screening with multiple isolates. The objective of this research is to investigate the genetic diversity among isolates from the pathogen S. trifoliorum from various European countries. We assessed diversity using a species identification test based on the sequence of the beta-tubulin gene, vegetative compatibility grouping and AFLP.     

Meiosis in NEUROSPORA CRASSA. I. the Isolation of Recessive Mutants Defective in the Production of Viable Ascospores

A scheme has been devised for efficient isolation of recessive meiotic mutants of Neurospora crassa. These mutants were detected by their reduced fertility or by the abortion of ascospores. Their isolation involved the selection and screening of the strains arising from ascospores disomic (n + 1) for linkage group I (LG I), which bears the mating-type locus. These strains are self-fertile heterokaryons that contain two types of haploid nuclei of opposite mating types (A + a). Selfings of these strains are homozygous for genes on all linkage groups except LGI and therefore allow the expression of recessive mutants with an altered sexual cycle. Using this selection procedure, three classes of mutants were detected. In one class, mutants had an early block in perithecial development (class I), and in another mutants had altered perithecia, but apparently unaltered fertility (class III). No recessive mutants were observed and all mutants tested (eight of class I and two of class III) were expressed only when used as the maternal parent. A third mutant class displayed normal production of perithecia, but defective formation of asci (class IIA), or black ascospores (class IIB). Four of 13 class IIA mutants were analyzed, and two of them [asc(DL131) and asc (DL400)] were definitely recessive analysis of 10 of 13 class IIB mutants disclosed six recessive, mutually complementing mutants: ase(DL95), asc(DL243), asc(DL711), asc(DL879), asc(DL917m) and asc(DL961). Mutants asc(DL95), asc(DL243) and the previously studied mei-1 mutant (Smith 1975) complemented one another in crosses, but did not recombine. These may be alleles of the same gene, or they may comprise a gene cluster.     

Taste Tests: Pizza and the Gastropolitical Laboratory in Mumbai

Abstract

This article is about experiments in taste. Focused on the cultural politics of pizza in Mumbai, it highlights the visceral work required to naturalize consumer choice as a catalyst of social futures in contemporary India. It emerges out of interviews and observations among food marketers and among customers and workers in a pizza restaurant. Guided by the concept of ‘experimentality’ elaborated in medical anthropology and science and technology studies, the article reframes definitions of food in experimental terms through two ethnographic registers. The first narrative focuses on the marketing of pizza, and explains how marketers turned their restaurants into laboratories of cosmopolitan cultivation. The second narrative shows how laboring bodies become enrolled in that laboratory, as young adults recruited from low-income neighborhoods come to work at a pizza restaurant. Such experiments blur the lines between evidence and enjoyment, and ultimately add up to a revaluation of public eating itself.     

A Comparison of Screening Techniques for Resistance to Verticillium Wilt in Alfalfa (Medicago sativa L.)

Three screening tests for resistance to Verticillium wilt in alfalfa using stem cuttings were compared including: stem infusion with a fungal culture filtrate (SIF), leaf injection with a spore inoculum (LII) and stem infusion with a spore inoculum (SII). The disease severity indice (DSI) determined by the three tests were very similar on average and were significantly correlated for a population of 142 plants regenerated from tissue culture. The DSIs determined by the LII test were also significantly correlated with the DSIs determined by the North American Standard test in a population of 20 Vertus (c.v.) plants. The results suggest that the assays using stem cuttings are effective for determining resistance to Verticillium wilt in alfalfa. Because they are nondestructive and quicker than the North American Standard test, these methods should find application in alfalfa breeding.     

Production of Cell Wall-Degrading Enzymes by the Phytopathogenic Fungus Sclerotinia sclerotiorum

The range of polysaccharide-degrading enzymes and glycosidases formed by the phytopathogenic fungus Sclerotinia sclerotiorum was monitored following growth on 16 carbohydrate substrates. Endo- and exoenzymes capable of degrading cellulosic, hemicellulosic, and pectinolytic polysaccharides were secreted. Pectinolytic activities were produced constitutively on all of the substrates tested. Cellulolytic enzymes were not induced in simple sugar (i.e., glucose or xylose) media. Polysaccharide growth substrates and cellulase inducers increased all of the enzyme activities tested. Gel filtration analysis revealed the appearance of new molecular forms of pectinase, β-xylosidase, and cellobiosidase during induction on pectin and carboxymethyl cellulose media.     

Ascospores in the type species of the genus Torulopsis

It has been found that the type species, T. colliculosa, of the form genus Torulopsis constitutes a perfect state forming ascospores. It is proposed that the same “Torulopsis Lodder et Kreger-Van Rij” based on a new type, be conserved against the earlier homonym “Torulopsis Berlese” which is based on the type T. rosea.