Increased protoplast yield from oat leaves and bean internodes by non-injurious mechanical perturbation

Summary A simple new technique has been developed to greatly increase the yield of protoplasts from plant organs without injury to the plant. Mechanical perturbation (MP) by non-stressful rubbing of oat leaf segments and bean internodes yielded ten to twenty times more viable protoplasts than did controls. The increase in protoplast yield due to MP is best manifested, if the organs are excised and transferred to the cellulytic enzymes immediately after MP is given to the intact organ. The enzymes begin digesting from the lower end of the bean internodes and proceed acropetally. Vacuum infiltration of control oat leaf segments for 15 min with enzyme solution resulted in increased yield but less than due to MP. Increased levels of calcium (10 mM) in the medium decreased the yield of protoplasts from both control and MP-treated plant organs. EGTA significantly increased the yield of protoplasts from control oat leaf segments and marginally over that found in the control bean internodes. Cycloheximide increased the yield of protoplasts from oat leaf segments but not from bean internodes. It is suggested that MP may increase the susceptibility of cell wall polymers to cellulytic enzymes by reducing calcium cross linking. MP is thus a tool for increasing the yield of protoplasts from plant organs without causing injury.Abbreviations CHI cycloheximide - EGTA ethyleneglycol-bis-(ß-aminoethyl ether)-N,N-tetraacetic acid - FDA fluorescein diacetate - MP mechanical perturbation     

Effect of presowing seed treatment with molybdenum and cobalt on growth,nitrogen and yield in bean (Phaseolus vulgaris L.)

Summary Bean (Phaseolus vulgaris L. Cv. Burpees Stringless) seeds were subjected to two cycles of presowing soaking and drying treatments with sodium molybdate and cobalt nitrite at 1 and 5 ppm concentrations used separately and also in combination. Sodium molybdate 2 ppm and cobalt nitrite 1 ppm used singly proved better than the remaining treatments with respect to nodulation, dry matter, nitrogen and yield. Combined treatment with sodium molybdate and cobalt nitrite did not produce additive effect on any parameter studied compared to their usage alone.     

南方菜豆花叶病毒外壳蛋白基因序列的分析

用重组DNA技术及序列分析法测定了南方菜豆花叶病毒RNA基因组3′端1,000个碱基的序列,以及由此序列推导出的整个外壳蛋白的氨基酸顺序,它与巳报导的基本上一致。介绍了用DNA的寡核苷酸水解混合物作为起始引物,以3′端不含PolyA尾巴且不能加上PolyA的病毒RNA作为模板合成互补DNA,及进一步无性繁殖此cDNA的方法。     

Examination of the influences of density pressure on the pattern of adult emergence with reference to the azuki bean weevil,Callosobruchus chinensis

Summary The effect of competition pressure on the sequential pattern of adult emergence was studied by using the experimental population of the azuki bean weevil. The density and pattern of emergence curves of the parental adult induced the changes in the shape of emergence curve of the progeny as well as the number of progeny individuals. In order to explain these changes, a simple mathematical model which has two independent variables, the population density and the capacity of environment, was introduced. The model generated basically similar patterns of emergene curves as observed in the present experiments. This means that the model displayed well the competition pressure which differentially acted upon early-born and late-born individuals in the populations being at different densities and which were differentially received by individual progeny according to the sequential distributions of parents. This different severity of competition pressure resulted in the difference of time when the pressure compelled influences on the sequential distribution of adult emergence. Therefore, early developed individuals may have good chance to survive, but the competition pressure works as a factor modifying their fundamental superiority, especially in the scramble type of competition. Intense crowding of the azuki bean weevil gave rise to the scramble type of competition in the relation between the numbers of adults emerged in two successive generations. When parental emergence concentrated in a short term and at high density, the competition becomes intense, resulting in the inferiority of early developed individuals, in the decrease in number at the next generation and in the increase of duration for emergence.     

Studies on interspecies competition between the azuki bean weevil and the southern cowpea weevil

Experimental studies was made on the interspecies competition between the azuki bean weevil, Callosobruchus chinensis and the southern cowpea weevil, C. macultatus. And the following results were obtained.

1. The reverse result of competition between the two species observed under the air-tight condition and the air-free one. That is, the population of the azuki bean weevils destroyedby that of the southern cowpea weevils under the former condition and vice versa under the latter. It is thought that such a reversal is due to the difference of sensibility of each species to the air-tight condition.
2. Under the air-free condition, change of the time interval of food-supplying had no effect on the result of competition within the limits of this experiment. The population of the southern cowpea weevils was always destroyed by that of the azuki bean weevils. But there was a certain degree of correlation between the duration of co-existence and the interval of food-supplying.
3. From the results, it is possible to say that by changing the degree of air-tightness, or the time interval of food-supplying, the co-existence period and the final result of competition can be changed.
4. The different mechanism of competition between two species in the present experiment from the experiments ofUtida (1952) andYoshida (1957) were discussed.
5. A difference in the mechanism of interspecies competition and intraspecies competition was expected from the level of total population numbers of two species and the individual weight of C. chinensis.

     

Distribution of diamine oxidase activity and polyamine pattern in bean and soybean seedlings at different stages of germination

Diamine oxidase (DAO, EC 1.4.3.6.) activity and polyamine content were measured in the shoot apex, leaves, epicotyl, cotyledons, hypocotyl and roots of light-grown bean ( Phaseolus vulgaris L. cv. Lingot) and soybean ( Glycine max L. cv. Sakai) seedlings at 3 different stages of germination (5, 8 and 14 days) as well as in embryos and cotyledons from soaked seeds. No DAO activity was detected in embryos and cotyledons of either plants. In bean seedlings DAO activity was only detectable in the shoot apex, primary leaves and cotyledons, while in soybean the activity was only detectable in the hypocotyl and roots. During seedling growth, in both plants, a different pattern of DAO activity was observed. In both species spermidine and spermine were the most abundant polyamines in embryos and cotyledons. Cadaverine, absent in bean, was only detected in soybean embryos. In the seedlings of both plants, increasing gradients of putrescine, spermidine and spermine from base to shoot apex were found. A high concentration of cadaverine was present in soybean hypocotyls and roots. A possible correlation between DAO activity and the endogenous content of the preferential substrate is discussed in relation to the possible involvement of the enzyme in regulating the cellular level of polyamines.     

Contagious egg-distribution on host pods and avoidance of pods received eggs in a wild bruchidKytorhinus sharpianus

The present study investigated the way a wild bruchid, Kytorhinus sharpianus, distributes its eggs on host pods. First, we counted the number of eggs per pod on the host plant (Sophola flavescens) in the field. The egg-distributions by the overwintered-generation adults in July and by the second-generation adults in mid October did not deviate from Poisson distribution because of small numbers of eggs laid. However, the egg-distribution in late August when many first-generation adults emerged was significantly contagious. The deposition of eggs was determined for three types of pods: (A) clean pods, (B) pods with eggs, and (C) those which eggs had been laid on but were removed from later. The numbers of eggs laid newly on three types of pods were significantly different; A>C>B, suggesting that an oviposition marker pheromone (O.M.P.) caused females to avoid pods on which eggs were deposited. Egg-shells reinforced that response. Monte Carlo simulations for the egg-distribution on pods predicted that even if females avoid pods with heavy egg-load through O.M.P., the egg-distribution becomes contagious when there is a large heterogeneity in quality of the pods.     

Identification of some major cytokinins in Phaseolus vulgaris and their distribution

The determination of the various endogenous cylokinins and their distribution among organs is important in understanding their role in growth and development in the intact plant. Cytokinins in young plants of Phaseolus vulgaris were purified by immunoaffinity chromatography and HPLC, and characterised by UV spectra. Zeatin nucleotide (zeatin riboside-5'-monophosphate) and isopentenyladenine nucleotide (isopentenyladenosne-5'-monopnosphate) were the most abundant cytokinins in all organs. Their identities were confirmed by GC-MS. The levels of zeatin, zeatin riboside, isopentenyladenine and isopentenyladenosine never exceeded 5% of the nucleotides, as assessed by a methodology that preserves cytokinin nucleotides. Three extraction methods were compared with qualitatively similar results, though differing in their suppression of nucleotidase activity. Cytokinin nucleotide levels were greater in the stems and petioles than in the roots and leaves on a per gram fresh weight basis, and were greater in the stems than in the other organs on a per plant basis. Levels of the zeatin and isopentenyladenine nucleotides were about equal in the stems and leaves, but in the petioles the zeatin nucleotide levels were about twice the level of isopentenyladenine nucleotide, while in the roots they were about half the isopentenyladenine nucleotide level. The importance of considering the cytokinin form is emphasised.     

Inheritance of resistance to potyviruses in Phaseolus vulgaris L. IV. Inheritance, linkage relations, and environmental effects on systemic resistance to four potyviruses

We have examined the genetics of systemic resistance in Phaseolus vulgaris to azuki bean mosaic virus (AzMV) and cowpea aphid-borne mosaic virus (CABMV) and the relationship of this resistance to a phenotypically similar resistance to watermelon mosaic virus (WMV) and soybean mosaic virus (SMV). In P. vulgaris cv Great Northern 1140 (GN1140), resistance to SMV and WMV has been attributed to the genes Smv and Wmv, respectively, which have been shown to segregate as a unit. Systemic resistance to AzMV is conferred by two incompletely dominant alleles, Azm1 and Azm2, at unlinked loci. At least three resistance alleles must be present at these two loci for systemic resistance to be expressed in the plant. Systemic resistance to CABMV in GN 1140 is conditioned by a dominant allele that has been designated Cam2. Under some environmental conditions, a recessive allele at an unlinked locus, cam3, also controls a resistant response to CABMV. Resistance to AzMV and CABMV does not assort independently from Wmv/Smv, but also does not consistently cosegregate, suggesting that perhaps in each case one of the factors involved in resistance is associated with Smv/Wmv.