Common bacterial blight, incited by Xanthomonas campestris pv. phaseoli (Smith) Dye (Xcp), is a serious disease of common beans (Phaseolus vulgaris L.). Three experiments were conducted twice in growth chambers at 26 ± 1C under short (10 hours light/14 hours dark) and long (16 hours light/8 hours dark) photoperiods to determine the influence of these photoperiods, flower bud removal, pod development, and pre- and post-inoculation photoperiods on the reaction of common beans to Xcp. In one test, `PC-50' (susceptible; S) flowered earlier and was more susceptible to Xcp under the short photoperiod than under the long photoperiod. BAC-6 (resistant; R) flowered at the same time under both photoperiods but developed rapid leaf chlorosis (RLC) (hypersensitive reaction) under long photoperiods. Flowering and disease reactions to Xcp by XAN-159 (R) were similar under both photoperiods. In a second test, daily removal of flower buds of `PC-50' decreased its susceptibility to Xcp under the short photoperiod. RLC of inoculated leaves of BAC-6 occurred during flowering and pod development under both photoperiods. XAN-159 expressed a high level of resistance to Xcp but showed RLC at later pod development stages. In a third test, the disease reaction of `PC-50' was affected by the particular photoperiod applied post-inoculation but was not influenced by the photoperiod applied before inoculation with Xcp. The implications of these results in breeding beans for resistance to Xcp are discussed.
Mohamed F. Mohamed and Dermot P. Coyne
Mohamed F. Mohamed, P. E. Read and D. P. Coyne
Regeneration in vitro from the embryonic axis in Phaseolus sp. has not been reported. Two embryo sizes, 0.3-0.4 mm and 0.6-0.7 mm long at 10-12 and 21 days after pollination, respectively, were excised from 4 P. vulgaris (P.v.) and 2 P. acutifolius (P.a.) genotypes. The embryonic leaves and radicale were removed, and 0.1-0.2 mm of the embryonic axis was cultured on Gamborg's B5 medium with 0, 5, 10 and 20μ MBA. The cultures were incubated in the dark at 25°C for 2 weeks followed by 1 week in continuous cool white light (25μ MS-1m2) before transferring to the second medium (0, 2μ MBA and 2μ MBA + 4μ MGA3). The tissues from the larger embryos initiated a single shoot without PGR in 30% of 1 P.v. explants and 30-60% in 2 P.a. The other 3 P.v. formed roots only. Multiple shoots were initiated in all P.v. (15-60%) and in 2 P.a. (60 and 70%) with 5 or 10μ MBA. The tissues from the smaller embryos had single shoots for all genotypes (30-60%) without PGR. Multiple shoots were initiated in 50-80% and 75-90% of the explants from P.v. and P.a., respectively, with 5 or 10μ MBA. Excess callus formed with 20μ MBA and regeneration decreased. After 3 weeks on the second medium, 6-8 shoot s/P. v. and up to 15-20 shoots/Pa. explants were observed.
Mohamed F. Mohamed, Paul E. Read and Dermot P. Coyne
Few studies on embryogenesis in common bean (Phaseolus vulgaris L.) have been reported and only the early stages of somatic embryogenesis were observed. Dry seeds from two common bean lines were germinated in darkness on L-6 medium containing 4% sucrose, 0.2 g casein hydrolysate /liter and 2.0 g phytagel /liter. The medium for seed germination was supplemented with 0, 2, 4 or 6μM forchlorfenuron (CPPU). Explants from cotyledonary leaves, petioles, hypocotyls and shoot apices were prepared from 14 day-old seedlings. Callus was derived from explant cultures incubated in darkness at 26C on the medium containing 4 μM 2,4-D and 1 μM Kinetin. The callus was transferred after 4 weeks into 125 ml Erlenmeyer flasks containing 50 ml liquid medium and placed on a gyrotary shaker (120 rpm) under cool-white light (12 μmol.m-2.s-1). The liquid medium was used with 2, 4 or 6 μM of 2,4-D alone or with zeatin supplements at relative concentrations of 0.25 and 0.5. Up to 200 somatic embryos from 40 to 50 mg callus inoculations were induced after 4 to 5 weeks. Callus derived from seedlings grown on CPPU-containing medium gave more repetitive somatic embryos. Cotyledonary stage embryos with clear bipolar structure were observed only from callus derived from seedlings grown on CPPU when transferred to suspension cultures containing 2,4-D and zeatin. All somatic embryos differentiated strong roots and some developed leaf-like structures on conversion medium.
Mohamed A. Shahba, Mohamed S. Abbas and Saad F. Alshammary
Understanding how mowing height and soil moisture influence drought resistance mechanisms may lead to better management of seashore paspalum. This research was conducted to evaluate the effect of mowing height and soil moisture replacement on drought tolerance strategies in three seashore paspalum cultivars. In a greenhouse, clear polyvinyl chloride (PVC) root tubes were placed in a black PVC sleeve with a bottom cap drilled with holes for drainage. Sod pieces (10 cm in diameter) of seashore paspalum (Paspalum vaginatum Swartz) cultivars Salam, Excalibur, and Adalayd were planted into these tubes after roots were trimmed. In a split-split experimental design, water regimes applied included control [100% of the total evapotranspiration (ET)] as well as 75%, 50%, and 25% of the total ET. Mowing heights were 45.0, 35, and 25 mm. Visual turf quality, maximum root extension (MRE), root length densities (RLD), total nonstructural carbohydrate content (TNC), shoot reducing sugar content (RSC), and proline content were determined. Turf quality decreased linearly with the decrease in irrigation water applied under the three mowing heights with higher slope at 25.0 mm than at either 35.0 or 45.0 mm. ‘Salam’ turf quality declined only to the unacceptable rating of 5.5 and 4.5 when mowed to 35 and 25 mm, respectively, whereas quality was 6.5 at the mowing height of 45 mm under the water regime of 25% of total ET. ‘Excalibur’ did not show acceptable turf quality at the 25% treatment, whereas ‘Adalayd’ did not show such quality at both 50% and 25% water regimes under all mowing heights. Regression analysis indicated a significant negative association between RLD and drought levels at all mowing heights and soil depths. In ‘Salam’, as drought levels increased from control to 25%, average RLD decreased by 76%, 75%, and 76% at 25-, 35-, and 45-mm mowing heights, respectively, at the top 30 cm of soil in the column. The change was 93%, 85%, and 83% at 25-, 35-, and 45-mm mowing heights, respectively, at the deeper soil (90 to 120 cm). In ‘Salam’, on average overall water regimes, MRE at 45 mm was ≈10% to 17% greater than that of 35-mm mowing height and 28% to 36% greater than that of 25-mm mowing height. The highest root mass (810 mg) was obtained when ‘Salam’ was mowed to 45 mm and subjected to the drought level of 50% of the total ET. The lowest root mass (320 mg) was obtained when ‘Salam’ was mowed to 25 mm and the water regime was not limiting. In ‘Salam’, as drought increased from control to 25% of the total ET, average TNC decreased by 43.5%, 26.0%, and 29.0% and the average TNC decrease in ‘Excalibur’ shoots was 48.0%, 30.0%, and 32.0%, whereas the decrease in ‘Adalayd’ was 51.3%, 42.3%, and 35.4% at 25-, 35-, and 45-mm mowing heights, respectively. As drought levels increased from control to 25% of the total ET, average RSC increased by 57.3%, 57.1%, and 53.0% in ‘Salam’ and by 59.4%, 57.0%, and 51.5% in ‘Excalibur’ and 61.2%, 58.1%, and 61.0% in ‘Adalayd’ at 25-, 35-, and 45-mm mowing height, respectively. When drought increased to 25%, average proline content in shoots increased by 435%, 432%, and 431% in ‘Salam’; 404%, 376%, and 324% in ‘Excalibur’; and 257%, 278%, and 302% in ‘Adalayd’, at 25-, 35-, and 45-mm mowing heights. The resistance of paspalum cultivars to moderate to high drought stress can be enhanced by increasing the mowing height that may be related to increased carbon fixation, which favors increased root production. Proline accumulation could add to the drought tolerance through osmoregulation or by acting as a carbon and nitrogen sink for stress recovery.
Mohamed F. Mohamed, Dermot P. Coyne and Paul E. Read
The leaf reaction of the Phaseolus vulgaris L. germplasm—UNECA (M6 mutant derived from the cultivar Chimbolito, Costa Rica), `Chimbolito', BAC-6 (Brazil), XAN-159 (Centro Internacional de Agricultura Tropical, Cali, Colombia), and `PC-50' (Domican Republic)—to Xanthomonas campestris pv. phaseoli strain V4S1 (Dominican Republic) were determined in two replicated trials conducted in a greenhouse in Lincoln, Neb. (Feb.–Mar. and July–Aug. 1993). `PC-50' and `Chimbolito' were susceptible to Xcp strain V4S1 in both tests. UNECA, BAC-6, and XAN-159 had similar levels of resistance to Xcp in the July to August trial. However, in the February to March trial, the resistance of UNECA was greater than that of BAC-6 but less than that of XAN-159.
Mohamed F. Mohamed, Dermot P. Coyne and Paul E. Read
Plant regeneration has been achieved in two common bean lines from pedicel-derived callus that was separated from the explant and maintained through successive subcultures. Callus was induced either on B5 or MS medium containing 2% sucrose and enriched with 0.5 or 1.0 mg thidiaznron/liter alone or plus various concentrations of indoleacetic acid. The presence of 0.07 or 0.14 g ascorbic acid/liter in the maintenance media prolonged the maintenance time. Up to 40 shoot primordia were observed in 4-week-old cultures obtained from 40 to 50 mg callus tissues on shoot-induction medium containing 1-mg benzyladenine/liter. These shoot primordia developed two to five excisable shoots (>0.5 cm) on medium with 0.1-mg BA/liter. A histological study confirmed the organogenic nature of regeneration from the callus tissues. The R2 line from a selected variant plant showed stable expression of increased plant height and earlier maturity. Chemical names used: ascorbic acid, N- (phenylmethyl)-1H-pnrin-6-amine [benzyl-adenine, BA], 1H-indole-3-acetic acid (IAA), N- phenyl-N'-1,2,3-thiadiazol-5-ylurea [thidiazuron, TDZ].
Mohamed F. Mohamed, Paul E. Read and Dermot P. Coyne
A new in vitro protocol was developed for multiple bud induction and plant regeneration from embryonic axis explants of four common bean (Phaseolus vulgaris L.) and two tepary bean (P. acutifolius A. Gray) lines. The explants were prepared from two embryo sizes, 3 to 4 mm and 5 to 7 mm, corresponding to pods collected after 15 and 25 days from flowering, respectively. The embryonic axis was cultured on Gamborg's B5 basal medium with 0, 5, 10, or 20 μm BA in combinations with 0, 1, or 2 μm NAA. The cultures were maintained at 24 to 25C under continuous light or incubated in darkness for 2 weeks followed by continuous light before transfer to the secondary B5 medium (0 or 2 μm BA or 2 μm BA plus 4 μm GA3). Adventitious roots or a single shoot with roots formed on the explants cultured on media without plant growth regulators. Multiple buds were induced on all BA media, but more were produced with 5 or 10 μm for most lines. Dark incubation greatly enhanced multiple bud initiation. Shoot buds were not produced on media containing NAA alone or in combinations with BA. On the secondary medium, six to eight shoots per explant for common bean and up to 20 shoots per explant from tepary bean were observed after 3 weeks. Mature, fertile plants were produced from these shoots. Chemical names used: benzyladenine (BA); 1-naphthaleneacetic acid (NAA); gibberellic acid (GA3).
Mohamed F. Mohamed, Paul E. Read and Dermot P. Coyne
Dry seeds from two lines of common bean (Phaseolus vulgaris L.) and one cultivar of faba bean (Vicia faba L.) were germinated on Murashige and Skoog (MS) medium containing B vitamins, 30 g sucrose/liter, and either 2.5, 5.0, or 7.5 μm benzyladenine (BA). Axenic seed cultures were grown at 22 to 24C in darkness and under continuous light from cool-white fluorescent tubes (40 μmol·m-2·s-1). Explant tissues were prepared from cotyledonary nodes (CN) and primary nodes (PN) of 14-day-old seedlings. Explants were cultured on corresponding seedling growth medium and maintained under continuous cool-white light (40 μmol·m-2·s-1). The percentages of CN and PN (in one line of common bean) explants that regenerated shoots and the number of shoots per explant (in all germplasm) were highest when nodal tissues were prepared from seedlings germinated in darkness. These responses were optimal on medium containing 5 μm BA during seedling growth and subsequent culture of explants. The number of shoots per explant was two to five times higher on explants cultured on medium with 0.25 to 1.0 μm forchlorfenuron (CPPU) or thidiazuron (TDZ) than on medium with 5 μm BA. Higher (2.5 and 5 μm) CPPU and TDZ concentrations inhibited shoot elongation and stimulated callus production. Histological analyses indicated that adventitious meristems formed 6 to 8 days after explant culture. Progenies from regenerated plants appeared similar to plants raised from the original seed stocks. Chemical names used: N- (phenylmethyl) -1 H- purin-6-amine (benzyladenine, BA); N- (2-chloro-4-pyridyl)-N'- phenylurea (forchlorfenuron, CPPU); N- phenyl -N' -1,2,3-thiadiazol-5-ylurea (thidiazuron, TDZ).
A. Dursun, D.P. Coyne, M.F. Mohamed and G. Jung
Common bacterial blight, incited by the bacterium Xanthomonas campestris pv. phaseoli (Xcp), is a serious disease of common beans [Phaseolus vulgaris (P. v.)]. Some tepary beans (P. acutifolius) are resistant (R) to Xcp and used to breed P. v. with R to Xcp. The objective was to determine the inheritance of the reaction to different strains of Xcp in crosses between susceptible (S) and R tepary lines. The parents, F2, and F3 populations from six tepary crosses involving 3 R × S, 1 R × moderately (M) R, and 2 R × R were inoculated with Xcp strains EK-11, LB-2, and SC-4A. Different single dominant genes controlled the reaction to different Xcp isolates in R × S crosses. Coupling linkage was detected between the genes controlling the reactions to each of the Xcp strains in the crosses NE #4B(s) × NE #19(R) and NE #4B(S) × CIAT-640005(R), except for NE #8A(MR) × NE #4B(S) with strains EK-11 and LB-2 and EK-11 and SC-4A. Transgressive segregation for S was observed in the F2 and F3 NE #8A × NE #8B(R), indicating that the parents possessed different genes for R. No segregation for reactions occurred n the F2 NE #8B × NE #19 and NE #19 × CIAT-640005, indicating that these parents possessed the same genes for R to the three strains.
Nader R. Abdelsalam, Rehab M. Awad, Hayssam M. Ali, Mohamed Z.M. Salem, Kamal F. Abdellatif and Mohamed S. Elshikh
Fig (Ficus carica L.) considers the original cultivated fruit trees and currently has become extinct. Such genetic resources should be identified, documented, and conserved. Morphology, pomology, and molecular markers are successful tools in assessing genetic diversity and classifying fig accessions. Twenty-one cultivated fig (F. carica L.) accessions were collected from Egypt and Libya. In Egypt, fig accessions are dispersed from Sinai in the east to El-Saloom in the west and from Alexandria in the north to Aswan in the south, whereas Libyan accessions were collected from Tubryq, Bengazi, and AlKufrah. Seventeen morphological, pomological, and fruit traits were used to characterize the fig accessions. Moreover, frozen young leaves were used to extract genomic DNA; 13 expressed sequence tag (EST) common fig primer pairs with 12 intersimple sequence repeat (ISSR)-anchored primers were used to detect the genetic diversity. Analysis of variance for fig accessions showed highly significant differences concerning morphological traits, i.e., the leaf length (centimeters) and width (centimeters) ranged from 5.4 and 6 cm to 23 and 23.5 cm, for Komesrey-El-Hammam, Abodey-Giza, and Black_Mission accessions, respectively. Also, fig accessions showed different shapes of leaf edge and fruits; they were categorized into four groups: straight, waved, zigzag, and serrated. The number of leaf lobes data ranged from one lobe for the ‘Green-yellow’, ‘Sultani Red Siwa’, and ‘Sultany Red Amria’ accessions to 10 lobes in the Aswany accession. The two-way hierarchical morphological cluster analysis distributed fig accessions into two main groups. The results detected high genetic diversity for the fig accessions that could be useful in the future breeding programs. Concerning molecular data, the EST markers showed highly polymorphism and informative (r = 0.61; 90.0%), with a total number of identified alleles of 78. We proved that a relatively greater number of alleles per locus characterizes the targeted loci among fig accessions, for which only one and two alleles per locus have been revealed, respectively, although ISSR showed a clear pattern and bands of the primers UBC807, UBC811, UBC812, UBC814, UBC815, UBC817, UBC818, and UBC823. In conclusion, a great range of variability was detected within the fig accessions. This diversification could enrich the genetic base of this genus, and more experiments are needed to reach its full potential.