Evaluation of annual wild <Emphasis Type="Italic">Cicer</Emphasis> species for drought and heat resistance under field conditions

About 90% of chickpea (Cicer arietinum L.) in the world is grown under rainfed conditions where it is subjected to drought and heat stress. Unlike the cultivated chickpea, annual wild Cicer species possess sources of resistance to multiple stress; annual wild Cicer species were therefore evaluated for resistance to drought and heat stress. Eight annual wild Cicer species (Cicer bijugum, C. chorassanicum, C. cuneatum, C. echinospermum, C. judaicum, C. pinnatifidum, C. reticulatum, and C. yamashitae) were compared with special checks, the cvs ICC 4958 and FLIP 87-59C (drought resistant) and ICCV 96029 (very early double-podded). ILC 3279 and 8617 as drought susceptible checks were sown after every 10 test lines. Yield losses due to drought and heat stress in some accessions and susceptible checks (ILC 3279 and ILC 8617) reached 100%. Accessions were evaluated for drought and heat resistance on a 1 (free from drought and heat damage)−9 (100% plant killed from drought and heat) visual scale. Four accessions of C. reticulatum and one accession of C. pinnatifidum were found to be as resistant to drought and heat stress (up to 41.8°C) as the best checks. C. reticulatum should be taken account in short term breeding programs since it can be crossed with the cultivated chickpea.     

Preliminary screening and selection for cold tolerance in annual wild <Emphasis Type="Italic">Cicer</Emphasis> species

The study was aimed to select cold tolerant accessions of annual wild Cicer species in the highlands in the west Mediterranean area of Turkey. A total of 43 accessions of eight annual wild Cicer species was screened for cold tolerance and compared with the best cold tolerant cultivars, ILC 8262 and FLIP 93-53C. After the sensitive check, ILC 533, was killed, accessions were evaluated using a 1–9 scale. Three accessions of Cicer bijugum, 2 accessions of Cicer reticulatum and 2 accessions of Cicer echinospermum were highly cold tolerant, while 3 accessions of C. bijugum, 9 accessions of C. reticulatum, 2 accessions of C. echinospermum and 1 accession of Cicer pinnatifidum were cold tolerant. All accessions of Cicer judaicum were sensitive to cold. Whereas Cicer chorassanicum and Cicer cuneatum were killed, Cicer yamashitae was recorded as highly sensitive to cold. The cold tolerant accessions of annual wild Cicer species were superior to the best cultivar as far as hardiness is concerned.     

Sources of resistance to cyst nematode in cultivated and wild Cicer species

Summary Among the nematodes infesting chickpea (Cicer arietinum L.) plants in Syria, cyst nematode (Heterodera ciceri Vovlas, Greco et Di Vito) is the most important. It is uneconomical to grow chickpea in fields infested with cyst nematode and to control this nematode with nematicide. Therefore, investigations were conducted at ICARDA, Syria from 1987 to 1991 to identify sources of resistance to cyst nematode in 7258 lines of C. arietinum and 102 lines of eight annual Cicer species including C. bijugum K.R. Rech. (13 lines), C. chorassanicum (Bge) M. Pop. (3 lines), C. cuneatum Hochst. ex Rich. (3 lines), C. echinospermum P.H. Davis (8 lines), C. judaicum Boiss. (18 lines), C. pinnatifidum Jaub. & Sp. (18 lines), C. reticulatum Ladiz. (36 lines), and C. yamashitae Kitamura (3 lines). All lines were grown in a greenhouse at 15–25°C in pots containing soil infested with 20 eggs of the nematode g^-1 soil. Nematode infestation was evaluated on a 0 to 5 scale based on number of females and cysts on roots. Resistance was found in one line of C. bijugum, six lines of C. pinnatifidum, and one line of C. reticulatum. No lines of C. arietinum, C. chorassanicum, C. cuneatum, C. echinospermum, C. judaicum, or C. yamashitae was resistant to cyst nematode. Plants with resistance have been recovered in the F₃ generation from crosses between the cultigen and C. reticulatum, indicating the possibility of transfer of gene(s) for resistance to cyst nematode from wild to cultivated Cicer species.Joint contribution from Istituto di Nematologia Agraria, ICARDA and ICRISAT (International Crops Research Institute for the Semi-Arid Tropics), Patancheru P.O., A.P. 502 324, India.     

Identification of resistant sources in Cicer species to Liriomyza cicerina

Summary Leaf miner (Liriomyza cicerina Rond.) causes considerable damage to chickpea (Cicer arietinum L.) in West Asia, North Africa and Southern Europe. Use of resistant cultivar is the economical method of control. Screening of over 7000 germplasm accessions did not result in identification of highly resistant accessions to this insect. Therefore, 200 accessions of eight wild Cicer species were evaluated in search of resistance to leaf miner between 1988 and 1991 at ICARDA, Syria. Accessions of Cicer species were screened under natural insect infestation in the field during spring (March–June), along with a susceptible-cum-indicator line. Two accessions of C. cuneatum (ILWC-40 and -187) and 10 accessions of C. judaicum (ILWC-44, -46, -56, -57, -58, -95, -103, -196, -206, and -207) were rated 2 on a 1–9 scale, where 1 = free from any damage and 9 = maximum damage. Another 18 lines of C. judaicum, four lines of C. pinnatidum and one line of C. reticulatum were rated 3 (resistant). Currently three species are incompatible in crossing with chickpea, but C. reticulatum is being used in breeding programme.Joint contribution from ICARDA and ICRISAT (Inter-national Crops Research Institute for the Semi-Arid Tropics), Patancheru P.O., AP 502 324, India.     

Endogenous Hormone Variations in Annual Wild Cicer Species

All processes that regulate of growth, differentiation and development and also stomatal movement are influenced by endogenous hormones in plants. Research related with endogenous hormones is known for cultivated chickpea (Cicer arietinum L.), while in wild chickpeas no data existed. In this study, C. bijugum K.H. Rech.; C. chorassanicum (Bge) M. Pop.; C. cuneatum Hochst. ex Rich.; C. echinospermum P.H. Davis; C. judaicum Boiss.; C. pinnatifidum Jaub. et Sp.; C. reticulatum Ladiz.; and C. yamashitae Kitamura were evaluated for variations in endogenous plant hormone concentrations; indole-3-acetic acid, zeatin, gibberellic acid, and abscisic acid in both leaf and pod. It was concluded that there was a great variation on endogenous plant hormones among the annual wild Cicer species. The data may support selection for various attributes.     

Identification of duplicates and fingerprinting of primary and secondary wild annual <Emphasis Type="Italic">Cicer</Emphasis> gene pools using AFLP markers

Wild annual Cicer gene pools contain valuable germplasm for chickpea improvement programs. Previous research showed that duplication might exist in accessions collected from these gene pools, which would hinder chickpea breeding and related research. AFLP (amplified fragment length polymorphism) markers were used to fingerprint the world collections of the primary and secondary gene pools including C. reticulatum Lad., C. bijugum K.H. Rech., C. judaicum Boiss. and C. pinnatifidum Jaub. et Sp. Duplicates were detected in a total of 24 accessions in both the gene pools, highlighting the necessity to fingerprint the germplasm. Genotypic difference was detected as gene pool specific, species specific and accession specific AFLP markers. These were developed into fingerprinting keys for accession identification between and within species and gene pools. Use of AFLP markers to detect duplicates and to identify accessions is a reliable method which will assist in the characterisation and use of wild annual Cicer germplasm in chickpea improvement programs. We recommend the procedure presented in this paper as a standard approach for the precise genetic identification and characterisation of future world collections of wild Cicer, to keep germplasm integrity and to benefit chickpea breeding and related research programs.     

Evaluation of perennial wild <Emphasis Type="Italic">Cicer</Emphasis> species for drought resistance

About 90% of chickpea (Cicer arietinum L.) in the world is grown under rainfed conditions where drought is one of the major constraints limiting its productivity. Unlike the cultivated chickpea, wild Cicer species possesses sources of resistance to multiple stresses; we therefore evaluated perennial wild Cicer species for resistance to drought. C. anatolicum, C. microphyllum, C. montbretii, C. oxydon and C. songaricum were compared with special checks; C. echinospermum, C. pinnatifidum and C. reticulatum and five cultivated chickpeas. After the cultivated chickpeas were killed, accessions were evaluated using a 1–5 scale, where 1 = highly drought resistant (no visible drought effect and full recovery after three successive wiltings) and 5 = highly drought susceptible (leaves and branches dried out, no recovery at all). All accessions of perennial wild Cicer species were significantly superior to those annual wild species and the cultivated chickpeas including the best drought tolerant chickpea, ICC 4958 under drought conditions. Perennial wild Cicer species did not only recover after wilting and drying out above ground level, they also tolerated high temperatures up to 41.8°C. But, they do not cross with the cultivated chickpeas. C. anatolicum should be taken account in long term breeding programs because it has closer affinities to the first crossability group than the others.     

Genetic relationships among perennial and annual Cicer species growing in Turkey as revealed by allozymes

Allozyme polymorphisms were used to assess genetic variation and relationships among ten Cicer species (annuals and perennials) growing in Turkey. Using seven enzyme systems, 12 putative scorable loci were detected and surveyed for polymorphism in an accession collection including wild and cultivated forms. Variation was generally low within accessions and species, but common between species. Cluster analysis based on the pairwise genetic distance coefficients among accessions and species using UPGMA revealed two species clusters; one includes three perennials (C. montbretii, C. isauricum and C. anatolicum) and the other contains six annuals (C. pinnatifidum, C. bijugum, C. judaicum, C. echinospermum, C. reticulatum and C. arietinum) and one perennial species (C.incisum). Grouping obtained in allozyme analysis appears to be consistent with the classification these species into three sections. However, contrary to relationships obtained in previous studies, three perennial species from section Polycicer were relatively distant to the group containing annuals. One perennial species, C. incisum from section Chamaecicer, clustered with annuals showing a close similarity. The grouping of six annual species was consistent with the previous reports of relationships. The relationships deduced between perennials and annuals appear to shed light on the evolution of annual habit from perennial habit.     

Evidence of rapid evolution and incipient speciation in Vicia sativa species complex based on nuclear and organellar RFLPs and PCR analysis

Seventy three accessions of the sevenVicia species belonging to Sativa speciescomplex were screened for nuclear and organellar restriction fragmentlength polymorphic (RFLP), and random amplified polymorphicDNA (RAPD) markers. Total genomic DNAs of 73 accessions wasrestricted with three enzymes, and the restriction fragments werehybridized to the wheat rDNA probe pTa71 (containing 18S, 5.8Sand 25S rDNA genes, and spacers), and faba bean probes Ver6-5 (entire intergenic spacer flanked by small part of25S and 18S fragments) and Ver 18-6 (part of thecoding region of the gene and internal transcribed spacers). InXbaI digests, 16 repeat unit length classes in24 combinations were identified. Digestion withEcoRI and DraI gave2–4 and 1–3 fragments, respectively, with detectablehybridization to the probes, indicating the existence of internalXbaI sites. All the accessions produced 3.5 EcoRI fragment arising fromcoding region of the repeat unit. Four hundred and eighteen RAPDmarkers among 45 accessions were identified with 14 arbitrary10-base primers. The percentage of polymorphic bands withinspecies ranged from 20 in V.angustifolia to 98% inV. nigra. Both RFLP andRAPD markers were unable to assess the relationships betweenaccessions within species as there was often much closer resemblancesbetween certain accessions of different species rather than betweenaccessions within each taxon. This analysis supports the view basedon morphological, cytogenetical and crossability data that it is notpossible to classify Sativa species complex into a small finitenumber of taxa which are clearly circumscribed, and that the complexrepresents a unique case of rapid evolution and incipient speciation.A study of chloroplast and mitochondrial RFLPs was undertaken toanalyze phylogeny through maternal lineage. Chloroplast DNArestriction fragment patterns, using 13 restriction endonucleases,revealed 92.6 to 99% homology between the seven species.Twelve enzyme-probe combinations yielded identical fragmentpatterns for all the seven species. The molecular sizes of thechloroplast DNAs obtained were similar (121.5–123.5), indicating that they had all lost one of theinverted repeats. Total DNAs digested with three restriction enzymesand hybridized to six heterologous probes of mitochondrial originyielded monomorphic bands in five enzyme—probe combinationsacross all the 73 accessions. In other combinations as well,40–66 accessions yielded monomorphic profiles. The smallvariation in the remaining accessions was not species-specificsince the same profiles were present in more than one species. Theseresults i) strongly suggest that the seven species within thecomplex share a common ancestor, or direct lineage and, ii)indicate that these species should be relegated to a rank, perhaps ofsubspecies, within V.sativa species complex.     

Perennial Wild Relatives of Chickpea as Potential Sources of Resistance to Helicoverpa armigera

The legume pod borer, Helicoverpa armigera (Hübn.), is one of the major constraints to chickpea production, and host plant resistance is an important component for the management of this pest. The levels of resistance in the cultivated chickpea are low to moderate, and therefore, we evaluated 17 accessions of perennial Cicer along with three cultivated chickpea genotypes for resistance to H. armigera. There was a significant reduction in both leaf feeding and larval weights when the larvae were fed on the leaves of Cicer microphyllum Benth. accessions ICC 17146, ICC 17236, ICC 17240, and ICC 17248. Relative resistance index based on leaf feeding, larval survival, and larval weight indicated that C. microphyllum accessions ICC 17146, ICC 17236, ICC 17234, ICC 17240, ICC 17243, and ICC 17248 were highly resistant to H. armigera. Under natural infestation, accessions belonging to C. microphyllum, C. canariense Santos Guerra et Lewis, and C. macracanthum M. Pop suffered a damage rating of <2.0 compared to 4.0 in C. judaicum Boiss. accession ICC 17148 (annual species) and 8.5–9.0 in the cultivated chickpeas (1 = <10% leaf area damaged, and, 9 = >80% leaf area damaged). There was considerable diversity in the accessions belonging to perennial wild species of chickpea, and these can be exploited to increase the levels and diversify the basis of resistance to H. armigera in the cultivated chickpea.     

Genetic diversity in a minicore collection of Cicer accessions using amplified fragment length polymorphism (AFLP)

This study was undertaken to establish the relationships among 44 chickpea (Cicer) accessions from a minicore collection including 30 cultivars, seven landraces from C. arietinum and seven accessions from C. reticulatum, C. echinospermum, and C. oxyodon species, all with economically important traits and tolerance to biotic and abiotic stresses, by using six amplified fragment length polymorphism (AFLP) primer combinations. The reaction products were resolved on MetaPhor agarose gel. A total of 64 clear and reproducible AFLP markers were recorded. Mean of polymorphism information content (PIC) values were calculated for each primer pair which ranged from 0.155 (EcoRI-ACC/MseI-CAG) to 0.270 (EcoRI-ACC/ MseI-CTG) with an average of 0.237. Analysis of molecular variance revealed that 90% of the total variance was due to differences within populations and 10% due to differences among populations. The results showed that the studied minicore collection is highly variable and could be used as the fundamental base in chickpea breeding programs.     

The extent of embryo and endosperm growth following interspecific hybridization between Cicer arietinum L. and related annual wild species

In vivo interspecific pollinations were performed and immature seed development investigated by histological methods in order to study crossability barrier(s) in Cicer L. species wide hybridization. Seven of the eight wild annual Cicer species, belonging to the secondary and tertiary gene pools, were used in reciprocal crosses with the cultivated chickpea. It was confirmed that the zygote was formed in all interspecific crosses. The embryos showed continued and retarded growth at different rate in various crosses, but eventually aborted at an early pro-embryo stage in all crosses except C. arietinum L. ×C. echinospermum Dav. Reciprocal cross differences were observed in early embryo growth rate and could have implications in obtaining hybrids. This study further emphasizes the necessity for developing appropriate and efficient in vitro procedures for rescuing immature globular hybrid pro-embryos, which will make the wild Cicer gene resources amenable to chickpea improvement.     

Molecular phylogeny of Chinese vegetable mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) based on the internal transcribed spacers (ITS) of nuclear ribosomal DNA

Sequence variation of nuclear internal transcribed spacer regions of ribosomal DNA (ITS1, 5.8S rRNA and ITS2) from Chinese vegetable mustards (AB-genome) and its putative parents Brassica rapa (the A-genome) and Brassica nigra (the B-genome) were used to investigate the molecular phylogeny and the probable evolutional pattern of this amphidiploid species that uniquely formed in China. Totally, 16 accessions of Chinese vegetable mustard those covering nearly all the diverse variations were included in this study, and together with three accessions of B. rapa and one accession of B. nigra. The results disclosed two strongly supported clades, one containing four accessions of vegetable mustard which have closer relationship with B-genome species “B.nigra” lineage and the other containing 12 accessions of B. juncea and three A-genome accessions. This classification was in disagreement with the evidence from chloroplast DNA, mitochondrial DNA, nuclear DNA restriction fragment length polymorphism (RFLP), which suggested that B. juncea was closely related to the A-genome type. For the incongruence, we speculated that the B. juncea crops derived from Chinese have evolved through different recombined events of the diploid morphutypes and evolved unidirectional concerted evolution. The traditional phenotypic classification of B. juncea was not wholly supported by ITS results, and hence the phylogenetic relationships among these subspecies need to be reconsidered on molecular level.     

RAPD and seed coat morphology variation in annual and perennial species of the genus Cicer L.

Molecular and morphological variation of six perennial and five annual species including domesticated chickpea, C. arietinum, were inferred on the basis of RAPD and S.E.M. seed coat features using three outgroup taxa (Lens ervoides, Lathyrus japonica and Pisum sativum). Of the 66 decamer arbitrary primers tested, eight primers revealed 87 informative fragments. Neighbor-joining cluster analysis using Jaccard's coefficient of similarity on the basis of polymorphic fragments indicated a narrow variation in C. arietinum and recognized two major clusters in the genus Cicer. The first one included four species of sect. Monocicer: C. echinospermum, C. arietinum, C. reticulatum and Iranian material of C. bijugum. The second cluster contained annual and perennial species belonging to sections Chamaecicer, Polycicer and Acanthocicer. The character state of morphological and ecological traits on the RAPD phenogram indicated no monophyletic incision. Our results suggested that the high genetic difference between annual and perennial species might be regarded as a rapid speciation of section Monocicer. Reconsideration of traditional classification of sections Polycicer and Acanthocicer is necessary. The Desi and Kabuli types of C. arietinum could not be separately grouped at the DNA level, and the low level of genetic variation of C. arietinum may result from a bottleneck during the domestication process.     

Delimitation of Amaranthus cruentus L. and Amaranthus caudatus L. using micromorphology and AFLP analysis: an application in germplasm identification

The ‘Morelos’ accessions of Amaranthus from Mexico demonstrate taxonomic ambiguity at the basic morphologic level. The main cause is the enormous morphological and genetic variation exhibited by the species in the genus. Although basic morphological criteria can be applied to herbarium specimens or germplasm collections for quick taxonomic identification, the morphological data alone can be misleading. To ascertain the taxonomic identity of the ‘Morelos’ accessions and their hypothesized species affiliation to Amaranthus caudatus or Amaranthus cruentus, we conducted a comparative analysis of phylogenetic relationships among these taxa/accessions using amplified fragment length polymorphism (AFLP) and micromorphology methods. Based on AFLP data, all the controversial ‘Morelos’ accessions can be consistently placed into a single A. cruentus species clade, which is clearly separated from the A. caudatus species clade. The AFLP-based phylogenetic relationship of ‘Morelos’ and delimitation of A. cruentus and A. caudatus are further supported by micromorphology, showing that the combination of these techniques can provide more reliable data for germplasm identification than each method used alone.     

Variation in resistance to the cabbage aphid ( Brevicoryne brassicae) between and within wild and cultivated Brassica species

Seven Brassica species were evaluated for their resistance to the cabbage aphid, Brevicoryne brassicae, in a series of field experiments. Four wild Brassica species, two 8 chromosome species with similarities to the B genome of Brassica nigra (Brassica fruticulosa and Brassica spinescens) and two 9 chromosome species containing the C genome (Brassica incana and Brassica villosa) were identified as possessing consistently high levels of antibiosis mediated resistance to B. brassicae. None of the species were shown to possess consistently high levels of antixenosis resistance. In more detailed glasshouse experiments one B-like genome species, B. fruticulosa, showed considerable variation between accessions collected from different sites for resistance to B. brassicae. In addition, individual accessions of one A genome species (Brassica rapa) and one C genome species (Brassica alboglabra) were shown to be highly variable in their resistance to B. brassicae, some plants of each accession being highly resistant and others very susceptible. The implications of the variability in resistance to B. brassicae within wild Brassica species for exploitation in Brassica breeding programmes are discussed.     

Molecular differentiation in Indian Citrus L. (Rutaceae) inferred from nrDNA ITS sequence analysis

Molecular differentiation in 24 accessions representing 19 taxa of Indian Citrus has been examined through sequence analysis of Internal Transcribed Spacer (ITS) region of nrDNA. Sequence length in the 24 accessions of Citrus taxa ranged from 512 to 665 bp (ITS1 & ITS2 partial and 5.8S complete sequence). The ITS sequences were very rich in G+C content ranging from 61.40 to 66.60% with an average of 64.2%. Genetic distance within Citrus group ranged from 0 to 13.4% with an average of 4.6%, showing moderate rate of nucleotide divergence. The phylogeny was inferred using the Maximum parsimony (MP) and Neighbor-Joining (NJ) methods. Both MP and NJ trees separated all the 24 accessions of Citrus into six distinct clusters. The disposition of all the accessions of Citrus in separate clusters in ITS-derived dendrograms was partly in accordance with the morpho-taxonomic affinities of the target taxa. This study supports the concept of Citrus medica (citron), C. reticulata (mandarin), and C. maxima (pummelo) as the basic species of the genus. However, ITS marker could not find any clear cut differentiation between subgenera Citrus and Papeda as proposed in Swingle’s Citrus classification system. The present study also supports the distinctiveness of C. indica (Indian wild orange), C. latipes (Khasi papeda) and C. hystrix (Melanesian papeda) as true species, besides elucidating the probable hybrid origin and relationships among the cultivated species/biotypes, such as Citrus ×aurantiifolia (sour lime) C. ×limon (lemon), C. ×taitensis (Indian rough lemon), C. limettioides (sweet lime), C. ×aurantium (including sour and sweet oranges and grapefruit), and other indigenous varieties of Indian origin: C. megaloxycarpa (sour pummelo), C. karna (karna orange), C. pseudolimon (Hill lemon), ‘Memang athur’, ‘Pummelo-lemon’ and ‘Kathairi nimbu’.     

Phylogenetic relationships of Cajanus cajan (L.) Millsp. (pigeonpea) and its wild relatives based on seed protein profiles

Total seed storage protein of 9 accessions of cultivated C. cajan and 10 wild Cajanus species was reported and compared by sodium dodecyl sulphate polyacrylamide gel electrophoresis. A considerable variation was detected among the protein profiles of different accessions of C. cajan while those of wild species were very specific and distinctly different from each other. Relative similarities between various taxa were estimated by Jaccard's similarity index and cluster analysis was performed to produce a UPGMA dendrogram. The clustering of 10 wild species and C. cajan more or less agrees with their sectional classification and available data based on morphological characteristics, crossability, genome pairing in hybrids and nuclear RFLPs. The species closest to C. cajan is C. cajanifolia although the accessions of C. cajan also share some bands present in the profiles of C. scarabaeoides, C. goensis, C. lineatus, C. acutifolius and C. volubilis. This points towards polyphyletic origin of the cultigen which has been discussed in this paper.     

Genetic Variation of Ethiopian Mustard (Brassica carinata A. Braun) Germplasm in Western Canada

Information on genetic diversity and genetic relationships among genotypes of Brassica carinata is currently limited. The objectives of this study were to evaluate patterns and levels of genetic diversity in B. carinata based on amplified fragment length polymorphisms (AFLP) as compared with Brassica juncea and Brassica nigra, and to evaluate agronomic and seed quality data for plants grown in the field in western Canada. A total of 296 AFLP bands were generated from four primer pair combinations and scored for presence/absence in 66, 20 and 7 accessions of B. carinata, B. juncea and B. nigra, respectively. B. carinata was less genetically diverse than the other two species. Differences in diversity were evident in the proportion of polymorphic loci within each species: 23, 35 and 50% for B. carinata, B. nigra and B. juncea, respectively. Pair-wise similarity measures based on the Jaccard coefficient were highest among accessions of B. carinata and showed the narrowest range: 0.911 (0.810–0.981) compared to B. nigra: 0.569 (0.438–0.660) and B. juncea: 0.715 (0.345–0.951). AFLP-based genetic distance information can be used by plant breeders to select diverse genotypes. AFLPs are also useful for fingerprinting cultivars and two primer pair combinations were sufficient to uniquely identify all the accessions of B. carinata. More variation among accessions was identified in the agronomic trial than had previously been described in studies of B. carinata in western Canada, but the data were too limited to draw conclusions regarding specific accessions. Overall, the findings were in agreement with other published work describing the favourable agronomic potential of this species.     

Phylogenetic relationships among species of Citrullus and the placement of C. rehmii De Winter as determined by Internal Transcribed Spacer (ITS) sequence heterogeneity

The internal transcribed spacer regions (ITS1 and ITS2) of the 18S-25S nuclear ribosomal DNA from the four recognized species of Citrullus (C. lanatus var. lanatus (Thunb.) Matsum. & Nakai., C. lanatus var. citroides (Bailey) Mansf., C. colocynthis (L.) Schrad., C. ecirrhosus Cogn., and C. rehmii De Winter) and Acanthosicycos naudinianus (Sond.) C. Jeffrey were amplified by PCR, and direct sequenced. Within the taxa examined, the length of ITS1 varied from 216 bp to 219 bp, and ITS2 varied from 239 bp to 249 bp. The average %CG content ranged from 59 to 64% and from 62 to 66% for ITS1 and ITS2, respectively. The greater length variation observed in ITS2 was primarily attributable to the occurrence of a (CC)_n microsatellite. Cladistic (PAUP) and phenetic (MEGA) analyses resulted in highly resolutive trees. ITS sequence analysis placed the recently described C. rehmii adjacent to the cultivated watermelon and supported the validity of the species classification of this taxa.