Heredity of seventeen isozyme loci in cassava (Manihot esculenta Crantz)

Summary Starch gel electrophoresis was used to assess isozyme polymorphism in two Manihot species. Crude extracts were obtained from leaves and pollen. Ten enzymes were examined for their polymorphism in a germplasm collection of 365 cultivated plus 109 wild accessions, mainly from Africa. The inheritance of these enzymes was examined using 13 intra and interspecific progenies. Seventeen polymorphic loci were found for the ten enzyme systems, with 59 alleles. All the markers showed disomic heredity and three linkage groups were identified.     

Characterization of resistance of cassava genotypes to bacterial blight by evaluation of leaf and systemic symptoms in relation to yield in different ecozones

Twenty-two improved and local cassava genotypes were evaluated for their bacterial blight symptom types in reaction to infection by Xanthomonas axonopodis pv. manihotis under field conditions in the forest, forest savanna transition and wet savanna zones of Togo. High genotype × environment interactions in development of each symptom type were observed. Combining data on environments and genotypes, spot, blight and wilt symptoms were positively correlated. Analysing genotype reactions across environments, indications for independent mechanisms of resistance on leaf and stem level, varying by genotype, were found. Genotypes Main27 with resistance to spot and blight symptoms and TMS4(2)1425 with resistance to wilt symptoms are recommended to breeders to introgress their resistance characteristics. Significant negative correlations were generally observed between blight and wilt symptom development and root yield across ecozones, with blight being more important under lower, and wilt under higher inoculum pressure. Genotypes TMS30572, CVTM4, TMS92/0429 and TMS91/02316 showed low spot, blight and wilt symptoms combined with high root yield across ecozones.     

RAPD-based genetic diversity study of fifty cassava (Manihot esculenta Crantz) genotypes

Fifty cassava clones were studied using RAPD technique. They included landraces from the Wenchi, Nkoranza, Dormaa Ahenkoro and Asonafo districts of the Brong Ahafo region of Ghana and three improved varieties. Genetic diversity of these genotypes was studied using four primers, OPK-01, OPR-02, OPR-09 and OPJ-14. A total of 41 different bands were detected. Levels of polymorphic fragments detected by the four primers ranged from 90% to 100%. By pooling bands from individual accessions together, mean number of fragments per accession per primer ranged from 5.50±1.04 for the Improved cultivars to 7.00±0.71 for populations of landraces from Dormaa. Mean frequencies of fragments not detected by the primers for the accessions were 0.524±0.12, 0.460±0.12, 0.561±0.12 and 0.523±0.12 for landraces from Wenchi, Nkoranza, Dormaa Ahenkro, Asonafo and the Improved varieties, respectively. The grand mean frequency of individuals showing fragments not present in populations was 0.522±0.10. Genetic diversity estimates ranged from 0.290 to 0.425 (mean 0.352±0.05) for primer OPK-01, 0.001 to 0.381 (mean 0.309±0.06) for primer OPR-02, 0.335 to 0.344 (mean 0.283±0.04) for primer OPR-09 and 0.152 to 0.352 (mean 0.261±0.07) for primer OPJ-14. Within the accessions mean gene diversity estimates were 0.316±0.03, 0.293±0.09, 0.331±0.02, 0.322±0.07 and 0.247±0.03 for accessions from Wenchi, Nkoranza, Dormaa Ahenkro, Asonafo districts and the Improved varieties, respectively. Interpopulational genetic divergence ranged from 0.069 to 0.203 (mean 0.119±0.04). Rate of nucleotide substitution among the landraces was 9.8 per cent per site per year, while that for the Improved varieties was 15 per cent. This revised version was published online in August 2006 with corrections to the Cover Date.     

Characterization and genetic distance analysis of cassava (Manihot esculenta Crantz) germplasm from Mozambique using RAPD fingerprinting

Twenty-eight cassava genotypes from Mozambique, along with seven genotypes from Angola, Madagascar, Nigeria, Togo, Columbia, and Thailand for comparison, were fingerprinted using random amplified polymorphic DNA (RAPD) analysis. The Mozambican material represented a wide range of landraces. A total of 311 scored RAPD loci were used to calculate genetic distances between the genotypes. This revealed an average genetic distance of 3.1% between all the germplasm. The average genetic distance between the Mozambiquen genotypes was 2.7%, whilst the seven accessions from the other countries showed an average distance of 3.4%. Neighbor-joining (NJ) method cluster analysis of the genetic distance yielded a tree that did not indicate a relationship between geographic distribution and genetic diversity. This revised version was published online in August 2006 with corrections to the Cover Date.     

Somatic embryogenesis from floral tissue of cassava (Manihot esculenta Crantz)

The aim of this study was to examine the embryogenic potential of floral material of the cassava cultivar MCOL 1505. Macerated immature inflorescences were found to be highly embryogenic, with almost 78% of the explants producing somatic embryos. Somatic embryos were also produced from whole male florets and half florets although at much lower rates. No regeneration was obtained from anther, microspore or floret wall tissue. Somatic embryos derived from immature inflorescences were regenerated via organogenesis and the plants derived from this process were assessed in terms of phenotype and ploidy level. If haploid plants could be produced by this method, this would have significant implications in assisting traditional cassava breeding, as this would allow homozygosity to be reached more rapidly. In a crop such as cassava, which is highly heterozygous in nature, the use of haploids in a breeding programme could considerably shorten the time taken to produce new desirable cultivars. This is the first report on plant regeneration through somatic embryogenesis from floral tissue of cassava. This revised version was published online in July 2006 with corrections to the Cover Date.     

Qualitative and quantitative evaluation of cassava bacterial blight resistance in F1 progeny of a cross between elite cassava clones

Deployment of resistant varieties is one major approach to controlling cassava bacterial blight (CBB), caused by Xanthomonas axonopodis pv. manihotis (Xam). To understand the genetic determinism of resistance to CBB, the use of reliable parameters measuring resistance is necessary. In order to test a relevant method for evaluation of quantitative resistance for mapping QTL (quantitative trait loci), the response of 150 F₁ individuals, inoculated with four different Xam strains (CIO-84, CIO-1, CIO-136 and CIO-295), was assessed under controlled conditions. We used two types of evaluations at different intervals after inoculation, one based on a scale of 0 to 5 and the second based on the determination of the bacterial population in the vascular system. Both evaluation types revealed interaction between strains and F₁ genotypes. Population values at 3 and 6 cm from the point of inoculation showed a high level of correlation. By performing an association analysis, at 7 and 15 days after inoculation, a significant positive correlation between both evaluation types was obtained. However, the disease rating at 30 days did not correlate with bacterial populations at either 7 or 15 days after inoculation, except for one strain, CIO-84. Evaluation of the bacterial population in stem tissues is time and labour consuming, consequently, for a rapid and reliable assessment of CBB resistance for QTL analysis, we strongly recommend evaluation based on the use of a symptom scale.     

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.     

Histological analysis of embryo-sac formation and detection of meiotic diplospory in cassava (Manihot esculenta Crantz)

Summary Embryo-sac formation in six cultivated Asian cassava clones and one unknown wild type, was histologically investigated to determine apomictic potential in the genus Manihot. All the 6 clones were found to possess one single sexual embryo-sac containing 6–8 nuclei at maturity. However, one clone, Rayong 3 possessed two functional embryo-sacs at 12 hours after controlled pollination (HACP). The larger embryo-sac was 6-nucleate (all 3 nuclei of the egg apparatus, 1 polar and 2 antipodal cells) and located towards the false micropylar or the nucellar beak region. The smaller embryo-sac also contained a total of 6 nuclei and approximately 10 m³ or 1/2 the volume of the former and located towards the chalazal pole. Percentage sexuality estimates indicated a 100% sexual reproduction in embryo-sacs of all the clones analysed. However, apomeiotic relationships indicate a low (0.3%) degree of meiotic diplosporous embryo-sac formation, thus suggesting facultative apomixis in cv. Rayong 3. This is the first embryological identification and evidence of apomeiosis and apomictic potential in cassava.Contribution from Plant Breeding Laboratory, Applied Genetics & Biotechnology Division, Faculty of Agriculture, Miyazaki University, No. 91.     

Inheritance of resistance to Xanthomonas campestris pv. translucens in triticale

Summary Three triticale lines, Siskiyou, M2A-Beagle, and OK 77842 have been reported to possess resistance to bacterial leaf streak caused by Xanthomonas campestris, pv. translucens (Xct.). The three resistant lines were crossed to susceptible lines and crossed with each other. F₂, BC₁-F₁, BC₂-F₁ plants were inoculated with a mixture of two Xct strains. The segregation data indicate the presence of a single dominant gene in each of the three resistant lines to bacterial leaf streak. These three genes are either the same or closely linked herein designated as Xct₁.     

Correlated resistance of cassava to mosaic and bacterial blight diseases

Summary The two most serious diseases of cassava (Manihot esculenta Crantz) are cassava mosaic disease (CMD) and cassava bacterial blight (CBB) (Xanthomonas manihotis Starr). Clone 58308, derived from the third backcross of the interspecific cross of cassava (M. esculenta) x ceara rubber (M. glaziovii), showed a high level of resistance to both diseases. Crosses of 58308 with several other clones which varied from susceptible to moderately susceptible to both diseases gave progenies with a significant genotypic correlation between resistance to both diseases (r=0.90), apparently due to linkage. The heritabilities of resistance to the diseases were estimated at 50–70% for CMD and 25–65% for CBB. Resistance to both diseases is assumed to be polygenic. The correlated response to selection for CMD and for CBB was estimated.     

Inheritance of resistance to Xanthomonas campestris pv phaseoli in tetary bean (Phaseolus acutifolius)

Summary The F1, F2 and F3 from two crosses within Phaseolus acutifolius were exposed to Xanthomonas campestris pv phaseoli to analyse the inheritance of resistance. The resistant parent, PI 319.443, gave a hypersensitive reaction in leaves and pods with small necrotic lesions. Based on the resistance of F1, the segregation in F2 and the reaction of F3 plants and lines, it is concluded that resistance in leaves and pods is governed by one dominant gene. Comparisons are made with the resistance to X. campetris in P. vulgaris.     

Identification of soybean strains resistant to Xanthomonas campestris pv. glycines

Summary Soybean germplasm was screened for resistance to bacterial pustule disease. The etiological agent, Xanthomonas campestris pv. glycines, was isolated from the leaves of field grown soybean in Maharashtra, India. The screening of soybean stocks was carried out by excised leaf inoculation method. A differential susceptibility to the pathogen was observed in the tested stocks. Two stocks P-4-2 and P-169-3 were found to be completely resistant to the pathogen and displayed an incompatible reaction. Four cultivars, EC-34160, Bragg, Kalitur and PK-472 displayed moderate resistance and the remaining stocks were susceptible to the attack of the pathogen. The stocks P-4-2 and P-169-3 remained resistant even to a high concentration of 10⁹ colony forming units (cfu)/ml of the pathogen.     

Resistance to common bacterial blight in Phaseolus vulgaris L. recombinant inbred lines under natural infection of Xanthomonas axonopodis pv. phaseoli

Among the main causes of poor yield in common beans are fungal, viral and bacterial diseases. Common bacterial blight, caused by Xanthomonas axonopodis pv. phaseoli (Xap), is one of the major bacterial diseases leading to significant losses in Brazil. Chemical control is ineffective, therefore, the use of resistant varieties becomes an interesting alternative. The objective of the present work was to evaluate disease resistance under natural infection of the pathogen in 109 recombinant inbred lines (F₇) of P. vulgaris originated from the cross HAB-52 (susceptible — snapbean) × BAC-6 (resistant — common bean) in two different environments, as well as to calculate genetic parameters to assist in the selection of promising materials to be used in the CBB resistance breeding program. The data of the genetic parameters were compared to those calculated for the F₃ generation originated from the same cross. The heritability results for DI (disease index) and VI (variation index) in F₃ were 26.85% and 0.26, respectively, whereas in F₇ they were 91.77% and 1.36, respectively. These results demonstrate a potential to be explored for this advanced population, that in the future, along with other pathogen variability studies and tests in other environments, may provide more information regarding a more precise evaluation of promising genotypes to be used in common bean breeding programs aiming to obtain CBB resistant varieties. This revised version was published online in July 2006 with corrections to the Cover Date.     

几个水稻不育系对白叶枯病的抗性

测定了８对不育系（Ａ）和保持系（Ｂ）以及一些杂交稻组合对６个小种的４０株白叶枯病菌的抗性，比较了不同类型不育系对６个小种群的抗性差异。结果表明，冈型、印尼水田谷型和大多数野败型不育系的抗性很差，ＢＴ型的抗性较高。菌株Ａｈ２８对冈４６Ａ和野败珍汕９７Ａ的致病力强。比较了６个恢复系（Ｃ）和８个杂交组合对不同小种的抗性程度。结果显示，杂交稻的抗病性主要受父本（恢复系）核基因的控制，但同时又受不育胞质的影     

Mating system in an experimental garden composed of cassava (Manihot esculenta Crantz) ethnovarieties

Crossing patterns were investigated in an experimental garden of ethnovarieties ofManihot esculenta (Euphorbiaceae) inPiracicaba, São Paulo, Brazil. A model of evolutionary dynamics for cassava presupposes genetic recombination by means of crossing within cassava gardens as a source that amplifies genetic diversity. Quantitative analysis of mating system parameters was performed using progeny arrays assayed for eight allozyme markers. The multilocus outcrossing rate (t _m)estimate (0.915±0.04)revealed that outcrossing was prevalent, but that a low level of self-pollination also occurred. The multilocus outcrossing rate ranged from 0.69 to 1.00 among eight varieties. The high value found for the outcrossing rate indicates that the ethnovarieties studied are preferentially allogamous. Genetic recombination occurred through crossing within the cassava garden, in agreement with an assumption of the model of evolutionary dynamics for this species.     

Efficient production of transgenic plants by Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz)

An efficient and reproducible method was developed for Agrobacterium-mediated transformation of embryogenic suspension cultures of cassava. LBA4404(pTOK233), containing the nptII, hph and gus marker genes, was used in the experiments. Chemical selection by means of kanamycin was used to establish 1037antibiotic resistant callus lines, of which 526 showed GUS expression. Of the 241 callus lines that were transferred to maturation medium 219formed somatic embryos. Thirty-seven of the 38 lines that were transferred to germination medium produced plants. GUS-positive plants could be obtained from 31 lines; in 14 of those lines 100% of the produced plants were GUS-positive, the remaining 17 lines yielded GUS-positive plants at an average of 72%. The transgenic nature of these plants was confirmed by Southern blot analysis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Nuclear DNA content and in vitro induced somatic polyploidization cassava (Manihot esculenta Crantz) breeding

Summary The diploid (2C) amount of DNA in cassava (Manihot esculenta Crantz) is 1.67 picograms (pg) per cell nucleus. This value corresponds to 772 mega-base pairs in the haploid genome. The size of the nuclear genome in cassava is very small in comparison with other Angiosperms. Flow cytometry techniques were used to screen ploidy levels in a large population of in vitro plantlets treated with colchicine and oryzalin (3,5-dinitro-N⁴,N-dipropylsulphate). Culture of axillary node cuttings for 48 hours in liquid medium supplemented with 2.5 to 5.0 mM colchicine in combination with 2% dimethyl sulfoxide (DMSO) resulted in a high frequency (23 to 42%) of non-chimeric tetraploids in the V₃ generation. Although mixoploidy may persist in as many as four cycles of vegetative propagation of node cuttings, solid (non-chimeric) tetraploids can be identified by flow cytometry among in vitro plantlets and then rapidly propagated for field testing. A somatic polyploidization system is proposed for implementation in cassava breeding programmes.Dedicated to the memory of the late Dr. Novak. Correspondence to M. van Duren     

Inheritance of resistance to black rot (Xanthomonas campestris pv. Campestris) in cauliflower (Brassica oleracea var. Botrytis)

Summary Black rot disease caused by Xanthomonas campestris pv. campestris is a limiting factor in the commercial production of the cauliflower crop. Crosses were attempted between SN 445, a mid season cultivar resistant to black rot and two highly susceptible commercial cultivars (Pusa Snowball-1 and K-1). Studies of the F₁'s, F₂'s and back crosses indicated that SN 445, carries a dominant gene imparting resistance to black rot.     

Heritability and gain from selection for quantitative resistance to Xanthomonas campestris pv. vesicatoria in Capsicum annuum L.

Summary Breeding for disease resistance in peppers (Capsicum spp.) to the bacterial spot pathogen (Xanthomonas campestris pv. vesicatoria (Doidge) Dye) has been based on either qualitative or quantitative evaluation methods. Quantitative evaluation of components of resistance, lesion number and lesion diameter, has been useful for determining quantitative resistance, but few breeders have applied these methods in routine selection programs. This study was aimed at determining the heritability and gain from selection for resistance to the bacterial spot pathogen based on three components of resistance. Random selections from a diverse intermated population of Capsicum annuum L. were self-pollinated for two generations to create S₁ and S₂ families. Thirty S₁ families, corresponding S₂ families and four homozygous check lines were evaluated. At forty-two days after seeding, two different leaves of each plant were inoculated by leaf infiltration with low concentrations (5×10³ colony forming units) of Group 2 (XCV PT, race 1) and 4 (XCV P, race 1) of the bacterium, respectively. After 15 days, lesion number cm^-2 and lesion diameter were measured. Total lesion area was calculated. Narrow-sense heritabilities for lesion number, lesion diameter, and total lesion area were 0, 43, and 31%, respectively, with Group 2, and 26, 43, and 33%, respectively, with Group 4. Actual S₂ gain from 20% selection pressure in the S₁ was approximately 50%, when selection towards resistance was based on total lesion area.