Identification of broad bean cultivars based on isozyme patterns

Summary With the increasing rate of new cultivar production for different crop plants, there is great need for methods of identifying each cultivar discriminatingly. Starch gel electrophoresis was employed to study the differences between the esterase and cathodal peroxidase isozyme patterns of 40 broad bean (Vicia faba L.) cultivars. A total of 10 and 17 medium to darkly stained bands were obtained for esterase and peroxidase systems, respectively. Bands from each enzyme system could be gropuped into three zones. Bands belonging to zone 1 of esterase (E₁) and zones 2 and 3 of peroxidase (P₂ and P₃) were quite distinct, stained intensely, and were especially useful for identification purposes.The differences in banding patterns among cultivars of the same origin were as great as those of cultivars of unrelated origin. A large proportion of the cultivars could be completely differentiated using both of the isozyme systems. There were no bands present in either enzyme system which were common to all cultivars.     

An electrophoretic survey of seedling isozymes in several Phaseolus species

Summary Starch-gel electrophoresis was used to study esterase, acid phosphatase, and peroxidase isozyme patterns in the primary leaves, stems, and roots of 13 species within the genus Phaseolus (P. acutifolius, P. adenanthus, P. angularis, P. atropurpureus, P. aureus, P. bracteatus, P. calcaratus, P. lathyroides, P. mungo, P. ricciardianus, domestic P. vulgaris, and wild P. coccineus and P. vulgaris). Most species showed unique banding patterns in each isozyme system. However, a close similarity in banding patterns was observed for the domestic P. vulgaris, wild P. vulgaris, and wild P. coccineus indicating close genetic relationships, possibly through species introgression or common descent.Great differences in isozyme patterns were found for different tissues from the same species in all systems indicating the high tissue specificity of isozymes. Upon compilation of bands from all the species, a very large number of isozymes were obtained in each system or tissue, making the electrophoretic isozyme technique of high potential in studies of isozyme variants in this genus.     

Species affinities between Cajanus cajan and some Atylosia species based on esterase isoenzymes

Summary Esterase isozymes were studied in seed extracts of Cajanus cajan and six Atylosia species by polyacrylamide gel electrophoresis and isoelectrofocusing. The isozyme patterns were stable and accession specific. Within the accessions of the Atylosia species, A. albicans and A. scarabaeoides showed three common bands indicating that they are more closely related to each other than to the other species. Of the accessions of Atylosia only A. cajanifolia shares the esterase isozyme of C. cajan and hence seems to be the closest wild relative of C. cajan.     

Protein electrophoresis as an aid to oat variety identification

Summary Ten cultivars of oats (Avena sativa L.) were investigated for their isozymes of esterase (E), leucine aminopeptidase (LAP), and anodal and cathodal peroxidase (APX and CPX).From repeated observations, a typical zymogram was prepared for each enzyme system, and the frequencies of the isozyme bands were scored for each variety separately. The results showed that: (1) most of the isozyme bands are very stable in their appearance on the gel: (2) each variety has a characteristic isozyme pattern; (3) 7 out of 10 oat cultivars showed no intravarietal isozyme variation. Some variation was found for the varieties Mesa, Rapida and Curt. This overall high level of homozygosity and homogeneity of these varieties for the isozyme bands makes this technique useful for varietal identification.     

我国棉花大丽轮枝菌(Verticillium dahliae)电泳图谱分析

对24株来自不同地区、作物的不同致病类型的大丽轮枝菌,进行了同工酶和可溶性蛋白电泳图谱分析。结果表明,24株大丽轮枝菌苹果酸脱氢酶酶谱具有种的特异性,酯酶酶谱具有属的特异性。谷氨酸脱氢酶、过氧化物酶、超氧化物歧化酶和可溶性蛋白电泳图谱与病菌的致病类型有关,可将24株大丽轮枝菌分为2群,群Ⅰ包括20个菌株,均属于中等致病力或弱致病力的非落叶型菌系;群Ⅱ包括4株菌,均属于强致病力的落叶型菌系。     

Isozyme variation between diploid and tetraploid cytotypes of Glycine tabacina (Labill.) Benth.

Summary Glycine tabacina (Labill.) Benth. is a wild perennial species related to the cultivated soybean, G. max (L.) Merr. It is composed of diploid (2n=40) and tetraploid (2n=80) cytotypes. Currently, to differentiate the cytotypes, plants are grown out in the greenhouse and chromosome counts made on pollen mother cells. It is a laborious and time consuming process. The objective of this study was to determine whether electrophoretic techniques could be utilized to separate the cytotypes. Electrophoretic examination of seven isozyme systems from seed of 67 G. tabacina accessions revealed banding patterns that could be used to differentiate between diploid and tetraploid cytotypes in the species. Among the tetraploid accessions, the number of bands observed were always greater than the diploids. Some tetraploid banding patterns consisted of bands similar to the diploid tabacina and/or additional bands previously identified in other Glycine species. The patterns of isozyme multiplicity and variation in the tetraploid tabacinas suggests more than one mode of origin for the tetraploids.     

Identification and polymorphism of cultivars and wild ecotypes of safflower based on isozyme patterns

Summary Acid phosphatase and cathodal peroxidase isozyme patterns were used to identify nine introduced and five local cultivars of common safflower (Carthamus tinctorius L.) and seven ecotypes of wild safflower (C. oxyacantha Bieb.). Nineteen and nine bands, respectively, were readily observed for the phosphatase and peroxidase systems in the extracts from seedling shoots. Results indicated that both systems could be used jointly for complete identification of all the 21 seed sources used.Band frequencies and polymorphic indices (PI) were determined from acid phosphatase patterns using at least 92 individual seedlings of five introduced and five local cultivars and three wild ecotypes. Band frequencies ranged from complete absence in some to total presence in other populations. The mean PI for the introduced, local and wild safflowers were found to be 0.069, 0.052 and 0.058, respectively.The banding patterns of either enzyme system were similar in some instances for the cultivated and wild safflowers and there was as much variation between two cultivars as between a cultivar and a wild ecotype, indicating close genetic relationships between C. oxyacantha and C. tinctorius.     

Isozyme electrophoregrams of sixteen enzymes in five tissues of cassava (Manihot esculenta Crantz) varieties

Summary Methodology, based on starch and polyacrylamide gel electrophoresis, was developed for determining isozyme electrophoregrams (patterns) of 16 enzymes of cassava (Manihot esculenta Grantz) varieties as potential genotype markers. Extracts of five different tissues (root, stem, leaf, petiole and bud) were examined. In general, the nodal portions of the shoots gave isozyme patterns with the largest number of bands. Petiole extracts gave similar results but bud extracts gave poor patterns. The limited number of varieties that were examined could be distinguished by sequential classification on the basis of the isozyme patterns of acid phosphatase, esterase, glutamate oxaloacetase transaminase and phosphoglucoisomerase.Joint publication of the Centro Internacional de Agricultura Tropical and the Department of Plant Science (No. 716), University of Manitoba. Presented in part at the 2nd International Symposium on Biochemical Approaches to Identification of Cultivars and Evaluation of Their Properties, 5–9 May, 1985, Braunschweig, West Germany.     

Characterization of cytoplasmic male sterility in Petunia hybrida. I. Localization,composition and activity of esterases

Summary Anthers of male fertile, cytosterile and restored male fertile clones of Petunia hybrida were compared for esterase activity and composition in subsequent stages of microsporogenesis. Three methods were applied (i) ultra-thin layer isoelectric focussing on polyacryl amide gels, (ii) quantitative spectrophotometrical assay, (iii) histochemical determination of total esterase activity associated to the azo-coupling method (Pearse, 1972).In male fertile and restorer idiotypes the isozyme patterns were quite similar. Both the number and intensity of bands increased gradually till the tetrad stage. In contrast, esterase activity in cytosterile anthers remained at a low level and hardy any new bands showed up in later stages. This unvariable, low activity level in cytosterile anthers was also found in the spectrophotometric assay. Histochemical determinations revealed that in male fertile anthers esterase activity is concentrated in the outer tapetal layer at late prophase and that it accumulates there till the early microspore stage. In male sterile anthers esterase accumulation in the tapetal cells stops at the moment that tapetal breakdown becomes visible. This suggests that differences in esterase activity and composition are an effect rather than a cause of the failing pollen formation.     

Diversity in the wild potato species Solanum chacoense Bitt.

Summary The morphological and isozyme variation was studied in 22 accessions of Solanum chacoense from Paraguay and Argentina. Clear geographic groups were identified through the use of multivariate analyses. S. chacoense from mountain sites in Argentina could be readily distinguished from plains forms from Paraguay, on the basis of several correlated morphological characters. Three isozyme systems, namely phosphoglucose isomerase (PGI), glutamate-oxaloacetate transaminase (GOT) and peroxidase (PRX) were studied using starch gel electrophoresis. The banding patterns indicated that for each isozyme there were several loci, which were polymorphic. A genetic interpretation of one of the PGI loci was made, and indices of genetic diversity and genetic identity calculated. Principal components analysis, cluster analysis and genetic diversity indicated a close relationship between the geographical groups. These results are discussed in the context of in situ genetic conservation.     

玉米过氧化物酶和酯酶同工酶与杂种产量的关系

以10个玉米自交系以及由它们组成的45个杂交种为材料,进行酶谱分析与单株粒重的测定。结果发现,玉米杂种 F_1的过氧化物酶同工酶 Px_4和酯酶同工酶 E_3的酶谱完全决定于其双亲。双亲酶谱相同,F_1与双亲一样;双亲酶谱不同,F_1出现双亲的互补酶谱。这两种酶的酶谱有差别的双亲杂交,能够产生高产的组合;这两种酶的酶谱没有差异的     

Chromosomal and genic structure of Brassicoraphanus related to seed fertility and the presentation of an instance of improvement of its fertility

Summary In order to elucidate the mechanism of low fertility of Brassicoraphanus, i.e., amphidiploids between Brassica japonica Sieb. and Raphanus sativus L., the chromosome number of 253 plants was studied during the 3rd–9th generations for their seed fertility. Meiotic irregularity showed no connection with degree of sterility. Brassicoraphanus consisted of euploids (2n=38), hyperploids (2n=39–43) and hypoploids (2n=34–37) with white or yellow flowers. The number of plants was highest in euploids and became lower as the chromosome number diverged from the euploid number. Further, seed fertility was highest and the range of its variation widest in euploids. The seed fertility of aneuploids became lower and its variation narrower in proportion to the number of chromosomes additional to or missing from the euploid number. Yellow-flowered plants were superior in seed fertility to white-flowered plants. Seed fertility of plants is primarily affected by their chromosome numbers and secondarily modified by genic effects. As a whole, seed fertility of Brassicoraphanus increased gradually and its variation widened with the advance of generations. This was explained mainly by the increase of balanced combinations of genes.     

Interspecific hybrids from cross incompatible relatives of sweetpotato

Summary Hybrids were obtained from Ipomoea interspecific crosses through ovule culture. The hybridity of the progeny obtained from I. triloba × IitI. trifida and (I. triloba × I. lacunosa) × I. batatas (4x) crosses was established by comparisons of floral morphology and analyses of peroxidase and esterase isozymes. The hybrids displayed the inflorescence type and sepal shape and texture of their male parents, while corolla size and anther and nectary color tended to be intermediate to their parents. The isozyme banding patterns of the hybrids contained bands present in the patterns of each of their parents. Pollen grain viability, measured by aceto-carmine stainability, was 44.1%, 92.3% and 82.4%, respectively, for the I. triloba × I. trifida hybrid and the (I. triloba × I. lacunosa) × I. batatas (4x) hybrids, H₁ and H₂. A controlled pollination study revealed that the I. triloba × I. trifida, and the (I. triloba × I. lacunosa) × I. batatas (4x) hybrids, H₁ and H₂ were partially self fertile with 6%, 70% and 13%, respectively, of the pollinated flowers producing viable seed. Success in backcrossing and sib-mating varied with the cross combination.     

Isozyme variation in Helianthus petiolaris and sunflower,H. annuus

Summary Helianthus petiolaris is a wild species used in genetic improvement of sunflower, as a donor of cytoplasmic male sterility and of genetic resistance to diseases. Isozyme variation for ADH, ACP, EST, GDH, LAP, PGI, PGD and SKDH in this species was studied using starch gel electrophoresis. The patterns thus obtained were compared with zymograms of inbred lines, hybrids and open pollinated varieties of H. annuus. The same alleles for EST and SKDH isozymes were found in both species, while ACP showed an allele that has not been found in sunflower. The rest of the isozyme systems showed both common alleles and characteristic ones for each species. ACP, GDH and PGD were monomorphic in H. petiolaris, while ADH and LAP were monomorphic in H. annuus. The isozyme markers obtained here could be useful in breeding programs involving interspecific crosses, and studies on introgression and on genetic variation in other populations of this wild species.     

Potential and limitations of isozymes for chromosomal localization of resistance genes against barley mild mosaic virus (BaMMV)

Summary To assess the possibilities offered by isozymes to locate resistance genes against barley mild mosaic virus (BaMMV), the isozyme patterns of 19 barley (Hordeum vulgare L.) genotypes carrying genes different from ym4 were determined. Of the 15 isozyme systems tested, only three were polymorphic, namely aconitate hydratase, esterase, phosphogluconate dehydrogenase, providing markers on four of the seven barley chromosomes. Studies of F₂ progenies derived from three crosses between resistant genotypes and susceptible varieties failed to reveal linkage between resistance genes and isozymes. Another goal of the experiment was to study the linkage relationships between ym4 and the esterase locus (Est1-Est2-Est4). Our estimates of the recombination rate between these two loci (3.41 and 8.32%) were in the range of those reported between these esterases and one of the resistance genes of the Chinese variety Mokusekko 3.     

Intergeneric crosses for the transfer of resistance to the beet cyst nematode from Raphanus sativus to Brassica napus

Summary The possibilities to transfer important traits and in particular resistance to the beet cyst nematode (Heterodera schachtii, abbrev. BCN) from Raphanus sativus to Brassica napus were investigated. For these studies B. napus, R. sativus, the bridging hybrid ×Brassicoraphanus (Raparadish) as well as offspring of the cross ×Brassicoraphanus (Raparadish) ×B. napus were used. Reciprocal crosses between B. napus and R. sativus were unsuccessful, also with the use of embryo rescue. Crosses between ×Brassicoraphanus as female parent and B. napus resulted in a large number of F₁ hybrids, whereas the reciprocal cross yielded mainly matromorphic plants. BC₁, BC₂ and BC₃ plants were obtained from backcrosses with B. napus, which was used as the male parent. F₁ hybrids and BC plants showed a large variation for morphology and male and female fertility. Cuttings of some F₁ and BC₁ plants, obtained from crosses involving resistant plants of ×Brassicoraphanus, were found to possess a level of resistance similar to that of the resistant parent. These results and indications for meiotic pairing between chromosomes of genome R with those of the genomes A and/or C suggest that introgression of the BCN-resistance of Raphanus into B. napus may be achieved.     

八个榆黄蘑菌株同功酶分析

为了探讨不同榆黄磨菌株之间的亲缘关系.采用垂直板聚丙烯酰胺凝胶电泳技术,利用酯酶同工酶和过氧化物同工酶酶谱的相似性和差异性,计算了8个榆黄蘑供试菌株间的Pearson相似系数;并采用类间平均法(Between-groups linkage)进行系统聚类,对供试菌株的亲缘关系进行分析.结果表明,所有供试菌株在2个同工酶系统中均有共同特征谱带,同时又体现了一定的多态性.菌株pc-5与其他各菌株遗传差异均较大;菌株pc-2与菌株pc-7的遗传差异很小,可能是源于同一种或为亲缘关系很近的变异类型;其余各菌株都有一定程度的差异性.     

应用过氧化物同工酶鉴定山西省马铃薯主栽品种的研究

为了快速、准确鉴定马铃薯品种,试验采用聚丙烯酰胺凝胶电泳分离马铃薯过氧化物同工酶(POD),并以酶带图谱作为鉴定依据,从而实现对马铃薯品种的鉴定。本文以14个不同品种的马铃薯的芽部、茎部、叶片为实验材料,分别提取其过氧化物同工酶进行聚丙烯酰胺凝胶电泳,分析比对同工酶的酶谱差异,进而确定不同品种的差异及其亲缘关系的远近。结果表明,马铃薯芽的过氧化物同工酶可以作为区分马铃薯品种的一个重要指标,14个品种中4、7号品种酶带相似,分别为晋薯11号和晋薯14号;6、9号品种酶带相似,分别为晋薯21号和晋薯18号;5、12号品种酶带相似,分别为同薯22号和同薯20号;13、14号品种酶带相似,品种分别为05-32-7和同薯23号;表明这些品种之间亲缘关系较近。     

The stabilization of chromosome numbers and the maintenance of euploidy in Brassicoraphanus

Summary To examine whether chromosome numbers of Brassicoraphanus (amphidiploids between Brassica japonica Sieb. and Raphanus sativus L.) are stable or not, the following four items were studied with some plants during the 2nd–11th generations: (1) chromosome numbers of open-pollinated progenies from eu-, hyper-, and hypoploids, (2) chromosome distribution at metaphase II in these plants, (3) frequency of euploids in relation to flower colour and generation, (4) seed fertility in eu-and aneuploids in relation to flower colour. In early generations, hyper-and hypoploids were frequently produced from euploids. In later generations, however, the chromosome number became less unstable. In euploids (2n=38), chromosome numbers at metaphase II showed some variation, and the mean frequency of the euploid chromosome number (n=19) was approximately 78%. This value was almost the same in white-and yellow-flowered plants through early and late generations. Nevertheless, yellow-flowered plants tended to produce euploids more frequently than did white-flowered ones. It is assumed that the difference in euploid productivity between yellow-and white-flowered plants is due to the difference in seed fertility between them. The progeny of each hypoploid showed higher chromosome numbers than their parents. The progeny of each hyperploid showed lower chromosome numbers than their parents: they were approaching to euploidy. This phenomenon, together with higher fertility of euploids and lower fertility of aneuploids, will favor the maintenance of euploidy of this strain.     

一粒小麦的染色体和同工酶分析

利用火焰干燥涂片法制备染色体玻片标本,观察到野生一粒小麦(T.boeo-ticum)和栽培一粒小麦(T.monococcum)的染色体组型基本相同。它们都具有着丝点带,但其中间带和末端带有些差别。在我们实验中它们的过氧化物同工酶及幼苗早期的酯酶同工酶酶谱也基本相似。因此野生一粒小麦和栽培一粒小麦可以归为一个一粒小麦种(T.monococcum),在小麦进化中,它们提供了 A 组染色体。