Selection for early flowering in progenies of interspecific crosses of ten species in the genus Dianthus

Summary Twenty-seven progenies of interspecific crosses, involving ten Dianthus spieces (D. caryophyllus, D. chinensis, D. superbus, D. barbatus, D. japonicus, D. brachyzonus, D. versicolor, D. trifasciculatus, D. serotinus and D. knappii) and some complex interspecific hybrids (Diantini genotypes and D. Allwoodii selections) were grown from seed under 8 hr days at two light intensities: 12 W/m² and 22 W/m². Observations on individual plants were made on vegetative development and date of flowering. After six months, 178 plants were selected from 23 seedling progenies and the resulting clones were compared under 22 W/m² and two photoperiods: 8 hrs and 24 hrs per day for a period of nearly six months. In the seedling stage, large differences were observed between progenies, e.g. in percentage generative plants (0–100%), plant length (3–78 cm) and plant fresh weight (0.5–68 g). Effects of light intensity were most pronounced in plant fresh weight (0.2–18 g in 12 W/m² and 0.5–68 g in 22 W/m²). Progenies of D. chinensis were the earliest to flower. Most progenies of crosses between D. superbus and D. barbatus segregated into flowering and non-flowering (rosetting) plants. In the clonal stage, most clones originating from non-flowering seedlings remained vegetative (rosetting), while all clones from flowering seedlings were either in flower or in bud within six months. Average dates of flowering of seedling progenies and of their clones were highly correlated, suggesting that selection on flowering date at the seedling stage is effective. High correlations were also found between the average flowering date of seedling progenies and the number and total weight of flowers harvested per plant in their clones within six months from planting. Overall effects of LD in the clonal stage were much less spectacular than the effects of light intensity in the seedling stage.     

Breeding for improvement of flowering attributes of winterhardy Rosa rugosa hybrids

Summary The purpose of this study was to determine at what stage of seedling development the flowering attributes in winterhardy R. rugosa hybrids could be assessed and if these attributes could be improved through breeding. The analysis of flowering was based on the duration of the flowering period and on amount of flowers produced.Most seedlings from rugosa hybrids could be assessed at an age of 3 years. The duration of the flowering period and flower productivity were not correlated. Several seedlings were improved in both attributes over the longer flowering and more floriferous parent.     

Evaluation of greenhouse inoculation techniques to screen sorghum for resistance to downy mildew

Summary Six inoculation techniques were compared for the artificial promotion of downy mildew (Peronosclerospora sorghi) in sorghum. These were (1) sprouted seeds incubated between sporulating infected leaves, (2) sprouted seeds depped in conidial suspension, (3) sprouted seeds sprayed with conidial suspension, (4) seedlings at plumule stage inoculated with drops of a conidial suspension, (5) seedlings at plumule stage sprayed with a conidial suspension, and (6) seedling showered with conidia falling from infected leaves. Seedlings at the one-leaf stage sprayed with a conidial suspension (6 × 10⁵ ml^-1) showed the highest systemic infection (100%) in the susceptible lines IS 643 and IS 18433. This technique is effective, repeatable, and allows the deposition of a conidial suspension as a fine mist on the entire seedling surface. In the greenhouse, the technique was used to test the downy mildew reaction of genotypes previously reported as resistant (< 5% incidence) in 3–4 years of field screenings. Of the 61 genotypes tested, 21 were free from downy mildew, 14 had less than 5% incidence, and the rest showed variable susceptible reactions. Therefore, the technique can be reliably and effectively used in the greenhouse to detect disease escapes and to indentify resistance.     

Screening for Fusarium resistance in seedling populations of Asiatic hybrid lily

A test to select Fusarium resistant seedlings of the Asiatic hybrid lily is described. Young seedlings of 28 populations, obtained from an incomplete diallel between eight parents with different levels of Fusarium resistance, were tested for resistance. Significant differences in Fusarium resistance between and within populations were detected. The average percentage of selected seedlings ranged from 34% in resistant × resistant crosses to 2% in susceptible × susceptible crosses. Although resistant descendants were obtained in susceptible × susceptible crosses, using at least one resistant parent produced higher percentages of resistant seedlings. The resistance level of the parents correlated highly with the general combining ability for Fusarium resistance based on the seedling test. For eight populations, seedlings selected for Fusarium resistance and non-tested (control) seedlings of the same cross were compared, after propagation, in a clonal test. Variation between and within populations, found at seedling level, was confirmed at clonal level. A positive selection response was found for all eight populations. In the seedling test, approximately 18% of the seedlings were selected as resistant of which 15% (2.7% of seedlings tested) appeared to be susceptible escapes. Comparison between selection at seedling level and at clonal level indicated that approximately 25% of the seedlings tested were missed (rejected resistant plants) in the seedling test. The practical use of a seedling test for Fusarium resistance in lily breeding programs is discussed.     

Light interception and radiation use efficiency in temperate quinoa (Chenopodium quinoa Willd.) cultivars

Sea level quinoas are grown at low altitudes in Central and Southern Chile. Both sensitivity to photoperiod and response to temperature largely determine quinoa adaptation, but crop biomass production must be quantified to evaluate agronomic performance. The objectives of this work are: (i) to characterize development effects on leaf area evolution for genotypes of sea level quinoa differing in cycle length, (ii) to quantify the extinction coefficient (k) for photosynthetically active radiation (PAR) and radiation use efficiency (RUE) from emergence up to the beginning of grain filling and (iii) to identify which crop attributes related to canopy architecture should be considered to improve biomass production. Four cultivars (NL-6, RU-5, CO-407 and Faro) were cropped in Pergamino (33°56′S, 60°35′W, 65 m a.s.l.), Argentina, at three densities (from 22 to 66 plants m⁻²) in two consecutive years under field conditions with adequate water and nutrient supply. Thermal time to first anthesis and maximum leaf number on the main stem were linearly correlated (r² = 0.87; p < 0.0001). Leaf area continued to increase during the flowering phase, notably in NL-6, the earliest genotype. There were significant differences in maximum plant leaf area between cultivars. Increasing density reduced plant leaf area but effects were comparatively small. Estimated k was 0.59 ± 0.02 across genotypes and was higher (p < 0.05) for 66 plants m⁻². Values for RUE changed as cumulative intercepted PAR (IPAR) increased; at initial stages of development RUE was 1.25 ± 0.09 g MJ IPAR⁻¹, but if cumulative IPAR was higher than 107.5 ± 10.4 MJ IPAR m⁻², RUE was 2.68 ± 0.15 g MJ IPAR⁻¹. That change occurred when leaf area index (LAI) and fraction of PAR intercepted were still low and ranged from 0.61 to 1.38 and from 0.33 to 0.51, respectively. No significant association was found with any developmental stage. Our results agreed to the notion that RUE variation during pre-anthesis phases is largely determined by LAI through its effect on radiation distribution within the canopy. Biomass production could be improved if periods of interception below 50% of incoming PAR were reduced to ensure high RUE. This seems to be possible in temperate areas both by the use of late genotypes with a higher number of leaves on the main stem and by early genotypes provided adequate plant density is chosen. Early increment in LAI and overlapping of the leaf area increase period with the flowering phase are desirable strategies for earliest genotypes to maximize yield.     

Variability and inheritance of flowering time and duration in Actinidia chinensis (kiwifruit)

We investigated the variability and inheritance of flowering in a seedling population of diploid Actinidia chinensis Planch. We found considerable variation in time to reach 50% flowering (D50) and flowering duration (FD). D50 was highly heritable (h ² > 0.85) suggesting that this trait would respond well to selection in this population, whereas FD was greatly affected by the environment (h ² < 0.20). The results also showed that the flowering times of male vines were more sensitive to the environment than those of female vines. However, significant specific combining ability (SCA) effects could be achieved by targeting specific bi-parental combinations for breeding and selection of flowering time. The diversity in flowering traits among males will facilitate the selection of male pollinizers that coincide in flowering time with present or future female cultivars. The possibility of breeding female cultivars with a short flowering period to improve the consistency of fruit maturity at harvest is discussed.     

低氮密植栽培对超级稻产量和氮素利用率的影响

为了研究低氮密植栽培对水稻分蘖发生及成穗率、干物质积累及其转化、氮素利用率和产量的影响,2012—2013年以超级稻Y两优1号为材料,在湖南长沙和海南澄迈进行了施氮量(75、150、225 kg N hm–2)与栽插密度(68、40、27、19穴m–2),每穴苗数(单、双、三本穴–1)与栽插密度(40、27、19、14穴m–2)的大田栽培试验。结果表明,在基本苗数相同或相近的条件下,减苗增密在齐穗期和成熟期的干物质量及产量分别比增苗减密高10.5%、5.2%和2.9%,有效穗数对产量的贡献最大,达到显著水平;在低氮密植条件下,有效分蘖期缩短6 d左右,分蘖成穗率、表观转化率、氮肥偏生产力和氮素籽粒生产效率分别提高10.9%、21.0%、150.6%和19.6%。在施氮量为75 kg N hm–2的密植(40~68穴m–2)条件下,齐穗期和成熟期的干物质量及长沙点产量分别比中、高氮(150~225 kg N hm–2)常规密度(19~27穴m–2)低3.2%、7.5%和1.2%,但差异不显著,而澄迈点产量在2012年和2013年分别比之低5.2%和高9.1%,且差异均达显著水平。在施氮量为150 kg N hm–2的密植条件下,成熟期干物质量比高氮常规密度低1.7%,但齐穗期干物质量和产量比高氮常规密度高10.3%和3.3%。因此,超级稻采用低氮密植栽培,在100~150 kg N hm–2和40穴m–2条件下提早了够苗期,增加了有效穗数,提高了分蘖成穗率和结实率,加之齐穗期适宜的干物质积累和较高的表观转化率,有利于高产的形成和氮肥利用率的提高。     

Accelerating and synchronizing germination of strawberry seeds by osmotic pre-treatments

Summary Pre-soaking strawberry seed (Fragaria ananassa Duch.) in osmotic solutions accelerated and partially synchronized their germination. Pre-soaking advanced 50% germination from about 4 weeks to 4 or 5 days. Two to three weeks pre-soaking in a mineral solution of -10⁶Pa nominal osmotic potential at about 20 C was satisfactory. The resulting synchronization of germination could allow seedlings to be selected for rate of growth by directly comparing seedling in bulk sowings.     

An evaluation of intergeneric hybridization of Fragaria spp. x Potentilla spp. as a means of haploid production

Summary Unidirectional intergeneric crosses using octoploid and hexaploid Fragaria species as a female parent and diploid and tetraploid Potentilla species as a male parent were performed, producing in toto 1345 fruits with an estimated 46089 seeds. The germinating seeds obtained by using P. anserina mostly perished at the cotyledon stage, a few developing into matroclinous octoploid seedlings. The seeds obtained by using P. fruiticosa produced two types of seedlings: matroclinous octoploids and genuine intergeneric hybrids. It is considered that the matroclinous octoploids resulted from contamination during the emasculation process. A great majority of the hybrid seedlings were slow in growth and development and perished before reaching the flowering stage. No haploids have been recovered. Possible reasons for absence of haploid genotypes in the progenies of these intergeneric crosses are considered. Among the seedlings of F. x ananassa cv. Tioga x P. fruiticosa, a seedling of an unusual phenotype was noticed, which upon chromosome determination, proved to be 2n=9x=63. This genotype displayed complete male and female sterility. The implication of the sterility of this seedling for bridging of the two genera is discussed.This research was performed as part of a cooperative project on genetic improvement of cultivated strawberry between the Fruit Laboratory, USDA, Beltsville and NJAES. Paper of the Journal Series, D-12102-16-83. New Jersey Agricultural Experiment Station, Rutgers University, New Brunswick, NJ 08903.     

Effect of temperature on gametophytic selection in a <Emphasis Type="Italic">Phalaenopsis</Emphasis> F<Subscript>1</Subscript> population

Gametophytic selection has potential to increase the efficiency of breeding for temperature tolerance. Here, we describe orchid seedlings after application of low and high temperatures during gametophytic development. In addition to phenotypic traits, amplified fragment length polymorphism (AFLP) markers were used to determine the genetic variability in seedlings. Two hybrid Phalaenopsis were cross-pollinated and exposed to 30°C day/25°C night for 3 days for a warm pollination or 15°C day/10°C night for 7 days as a cold pollination treatment. The plants were returned to the greenhouse after pollination and green capsules were collected after 150 days. Protocorms obtained from these treatments were evaluated 72 days after initial plating for germination and size on a thermogradient table ranging from 10 to 30°C. Seedlings were then evaluated 1 year after initial plating. The mean number of roots per seedling (4.2) was greater for plantlets that derived from the cold pollination treatment compared to those from warm pollination (3.6). Weight of the seedlings, number of roots and the average root length were significantly affected by the interaction between pollination treatment and germination temperature. The weight, number of leaves, and average root length were significantly affected by the interaction between pollination treatment and incubator/growth chamber. The results indicated that seedlings derived from warm pollination were more vigorous under warm growing conditions and those derived from cold pollination were more vigorous under cold growing conditions. Genetic variation among 16 F₁ seedlings randomly selected from various temperature treatments was analyzed. A dendrogram based on 651 loci resulted in three major groups and one subgroup. The groups and subgroup revealed common selection pressure during the gametophytic stage. The AFLP data support genetic differentiation of Phalaenopsis hybrids pollinated under different temperatures.     

Radiation use efficiency and leaf photosynthesis of sweet corn in response to phosphorus in a cool temperate environment

Phosphorus (P) deficiency has been shown to decrease accumulated intercepted solar radiation (RI_cum) for sweet corn and in this paper the effects on radiation use efficiency (RUE) and leaf photosynthetic rate at 2000 μmol m⁻² s⁻¹ PPFD (P₂₀₀₀) are examined. Data from two consecutive field experiments on a low P site at Lincoln, New Zealand, were analyzed. In the first experiment (2001/2002) 0, 50, 100, 150 or 200 kg P ha⁻¹ was applied to sweet corn followed by an additional 0, 0, 10, 20 or 40 kg P ha⁻¹ in 2002/2003 applied to the same plots. Thus, total P applications were 0, 50, 110, 170, or 240 kg P ha⁻¹.There were no differences in RUE between P treatments but RUE changed with crop ontogeny. The RUE was 0.66 g MJ⁻¹ before each crop had 10 fully expanded leaves and RUE was 1.34 g MJ⁻¹ after this. The cause of this difference between development stages was unclear, but it was not related to air temperature. In contrast to RUE, there were clear differences in P₂₀₀₀ due to differences in specific leaf phosphorus (SLP). At a SLP of 0.12 g P m⁻² or greater, P₂₀₀₀ was a constant value of 34 μmol CO₂ m⁻² s⁻¹. When SLP was less than 0.12 g P m⁻² P₂₀₀₀ was reduced. The differences in P₂₀₀₀ between P treatments occurred when the plants were young (≤10 fully expanded leaves) and after this, SLP of all leaves exceeded 0.12 g P m⁻² and there were no differences in P₂₀₀₀. These early leaves were unimportant in determining crop RUE but are likely to have been important in establishing the hierarchies that led to the changes in RI_cum reported previously. These results demonstrate the importance of an adequate supply of P early in crop growth.     

Intergeneric hybrid of two rare and endangered orchids, <Emphasis Type="Italic">Renanthera imschootiana</Emphasis> Rolfe and <Emphasis Type="Italic">Vanda coerulea</Emphasis> Griff. ex L. (Orchidaceae): Synthesis and characterization

Renanthera imschootiana and Vanda coerulea are two rare and endangered orchid species featuring in IUCN Red List of Threatened Plants as well as in Red Data Book of Indian Plants, and are renowned for their unique ornamental traits. Renanthera imschootiana and V. coerulea have different flowering periods and by taking opportunity of off-season flowering of V. coerulea which happened during the flowering season of R. imschootiana, the hybridization was effected by hand-pollination. Viable pollinia were present in the off-season flowers of V. coerulea and one out of four cross was successful when R. imschootiana was taken as female parent. Reciprocal cross was unsuccessful. The resulting immature hybrid embryos were germinated in vitro on Vacin and Went medium supplemented with 15% v/v coconut water. Best seedling growth was observed on half-strength Murashige and Skoog medium devoid of any plant growth regulators while the transplanted seedlings grew best on brick chips:charcoal pieces (2:1) potting mix in community earthen trays. First flowering of the hybrid seedlings happened four years ten months after transfer to ex vitro environment. RAPD markers generated by the decamer primer OPA1 convincingly confirmed the hybridity. Registration of the hybrid was made with the Royal Horticultural Society with the grex Renantanda Kebisana Shija.     

Rooting of bean leaves and use in germplasm evaluation for common bacterial blight resistance

Summary A simple protocol for leaf rooting in beans (Phaseolus vulgaris L.) was developed and used to investigate the reaction of Xanthomonas campestris pv. phaseoli (Xcp), causal pathogen of common bacterial blight disease, in detached versus attached bean leaves. Trifoliate leaves of different sizes (one-third, two-thirds, and fully expanded), either with or without the pulvinus attached to the petioles, were excised from 20 day-old plants of six bean cultivars/lines. Leaf cuttings were cultured in potting medium and then incubated for 5 to 10 days under transparent polyethylene plastic cover in the greenhouse. Roots were readily initiated along the petioles of the leaf cuttings, whether the pulvinus was present or absent. All leaves which were two-thirds expanded and fully expanded developed roots 5 to 7 days after culture. Eighty to 90 percent of the leaves which were one-third expanded formed roots 8 to 10 days after incubation. Laminae of the rooted leaf cuttings were viable and green during the 2 to 3 months period in culture after removing the plastic cover. The common bacterial blight reactions were similar for inoculated attached leaves, detached rooted leaves (inoculated either after or prior to rooting), and moistened detached leaves incubated without rooting. The latter were only usable for evaluation of the Xcp reaction in growth chamber experiments but not under greenhouse conditions. The rooted leaves would be useful for screening bean lines for multiple disease resistance, especially if the pathogens require different environments for disease expression.Abbreviations CBB Common Bacterial Blight - Xcp Xanthomonas campestris pv. phaseoli (Smith) Dye     

Stimulation of Rice (Oryza sativa L.) Seedling Vigour by Smoke-water and Butenolide

The newly identified smoke‐compound butenolide (3‐methyl‐2H‐furo[2,3‐c]pyran‐2‐one) can improve seedling vigour. This study highlights the effects of smoke‐water and butenolide on shoot and root elongation of a local rice variety. Smoke‐water (1 : 500) and butenolide (10^?8 m ) treatments significantly increased shoot length. A low concentration of butenolide (10^?10 m ) promoted maximum root length and seedling weight, which were significantly different from the control. Butenolide‐treated (10^?8 m ) seedlings had a significantly greater number of lateral roots than untreated seedlings. The vigour index of smoke‐water (1 : 500) and butenolide‐treated (10^?8 m ) rice seeds was significantly higher than that of untreated seeds. These findings suggest the possible use of butenolide for promoting vigorous seedlings in rice.     

Cotyledon angle as a predictor of growth rate of cucumber plant

Summary Experiments conducted in the greenhouse and in the field indicated that plants produced from seedlings of Cucumis sativus L. which displayed their cotyledons in an upward position grew faster than plants which displayed their cotyledons in a downward position. Cotyledon angle was not a function of seed size nor seedling weight at 10 days after planting. An increase in plant height, node number, and fresh weight at later growth stages was associated with an upward cotyledon angle and persisted until time of harvest. Anthesis tended to occur later in plants having a downward cotyledon angle. The number of pistillate flowers was greater in plants with an upward cotyledon angle compared to those with a downward cotyledon angle. Differences in yield (fruit number) were not associated with cotyledon angle.Paper No. 5432 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, NC. The use of trade names in this publication does not imply endorsement by the North Carolina Agricultural Experiment Station of products named, nor criticism of similar ones not mentioned.     

Variation in the photoperiodic response on several characters in soybean,Glycine max (L.) Merrill

Summary The photoperiodic response on 12 characters in 40 soybean, Glycine max (L.) Merrill, cultivars were investigated using 10-hour (short day) and 16-hour (long day) photoperiods. Seventeen cultivars showed no significant difference between photoperiods for all 12 characters. Seven cultivars showed significant difference for only one trait. Phenotypic changes in 17 cultivars may be due to factors other than photoperiod. Number of pods per plant, days to maturity, and number of nodes per plant were highly correlated with yield per plant in both photoperiods. Invariably, the cultivars which had significant differences in the numbers of flowers produced per plant between the two photoperiods also had significant differences in the numbers of pods per plant and yield. First node to flower, plant height at flowering, days to flowering, nodes at flowering, and the 100-seed weight were least influenced by the photoperiods in most of the cultivars, while the height at maturity, number of nodes at maturity, number of days to maturity, and yield were most influenced.AVRDC Journal paper 46 (78–88).     

Association between growth habit and persistence in red clover

Summary Three double-cut red clover (Trifolium pratense L.) varieties: Prosper I, Florex, and Tristan each with two different seedlots were space-planted in a field. The total number of plants established for each seedlot was about 900. In the seedling year, plants were classified into five growth types according to rosette development and flower production. In the fall of the third year 14% of the rosette-forming, non-flowering Type 1 plants were alive; 8–11% of the intermediate types survived; and only 2% of the nonrosette-forming, profusely flowering Type 5 plants remained growing. The non-flowering class (Types 1 and 2) persisted better than the flowering class (Types 3, 4, and 5). This difference in persistence was similar for all three varieties regardless of seed source. The two flowering classes, in the production years, had the same proportion of the most vigorous plants. The most vigorous plants of the non-flowering class had the same frequency distribution of seed yield as those of the non-flowering class. In conclusion, the flowering response can be used as a selection criterion for persistent plants; and plants not flowering in the seedling year can attain a forage and seed yield as high as flowering plants in the production years. Screening for non-flowering plants can be carried out at the seedling stage in the greenhouse before seedlings are transplanted to the field.Contribution No. 520, from the Charlottetown Research Station, Agriculture Canada, Prince Edward Island, Canada.     

Identification of growth stage dependent expression of partial resistance of barley to powdery mildew

Summary Genotypic differences for growth stage dependent expression of partial resistance to barley powdery mildew have been identified on the basis of two components of resistance in the glasshouse and the pattern of epidemic development in the field.Differences for infection frequency and proportion of sporulating colonies were highly significant between the 16 genotypes investigated at four stages of plant development in the glasshouse. Both resistance components were significantly correlated mutually (r=0.73 to 0.86) and with the infection level on leaves developed at comparable growth stages in the field (r=0.52 to 0.73). The infection level of seedlings in the glasshouse was significantly correlated with the infection level of the leaves of the first until the third node in the field (r=0.70 to 0.73). Adult plant resistance was predominantly expressed at the uppermost leaf. A clear genotype × growth stage interaction was apparent for infection level; some genotypes showed partial resistance predominantly at the seedling stage and others predominantly at the adult plant stage.Abbreviations IF Infection Frequency, number of colonies per cm² leaf area, glasshouse experiment - IL Infection Level, number of colonies per cm² leaf area, field experiment - PSC Proportion Sporulating Colonies from total number of colonies, glasshouse experiment     

Analysis of differences in growth of cucumber genotypes under low light conditions in relation to night temperature

Summary Growth of four breeding lines and two cultivars of slicing cucumber with different growth at low temperature and light intensity was analysed in relation to night temperature. Plants were grown from 17 days after sowing at 20°C day temperature and 20, 15 or 10°C night temperature (NT), a light intensity of 24 W/m² visible radiation and a daylength of 8 hours. Data from six successive harvests, ending 47 days after sowing, were analysed according to a MANOVA procedure.RGR, NAR, LAR, SLA and LWR generally decreased with time. Ln W and RGR were largely similar at 20 and 15°C NT, but reduced at 10°C NT. LAR and SLA decreased at each lower NT, whereas LWR increased. NAR was only marginally affected by NT. Significant genotypic differences occurred for 1n W, LAR, SLA, LWR, stem length and percentage of dry matter in leaves and stems at all NT's and for RGR and NAR only at 20°C. Genotypic differences for 1n W could not be explained by differences for RGR, but by differences for initial plant weight.Significant genotype × night temperature interactions were present for LWR. Breeding lines increased LWR in response to lower NT, while that of cultivars remained fairly constant. Implications for breeding are discussed.Abbreviations and definitions L plant length - LA leaf area - W dry weight of the shoot - RGR relative growth rate (the increase in W per unit of time and unit of W) - NAR net assimilation rate (the increase in W per unit of time and unit of LA) - LAR leaf area ratio (the leaf area at time t per unit of W) - SLA specific leaf area (the leaf area at time t per unit of leaf weight) - LWR leaf weight ratio (the leaf weight at time t per unit of W) - RGR NAR × LAR - LAR SLA × LWR - MANOVA multivariate analysis of variance     

Factors affecting in vitro flowering and fruiting of green pea (Pisum sativum L.)

Multiple shoots were efficiently regenerated from cotyledonary node and shoot tip explants of Pisum sativum within 15 days on MS medium containing B₅ vitamins and supplelmented with 2.0 mgl^-1 6-benzylaminopurine. The elongated shoots produced on the same medium were excised and transferred to MS medium containing half strength ammonium nitrate (8.25 gml^-1) and supplemented with auxins (indole-3-butyric acid or naphthalene acetic acid) either alone or in combinations with gibberellic acid. Rooting and flowering were observed on the 7th and 15th day after their transfer to rooting medium. The flowers self-fertilised in vitro and produced mature pods within 25 days of rooting. These seeds were germinable both in vitro and in vivo. In vitro seeds sown in pots under field conditions developed into flowering plants, and subsequently produced pods with viable seeds. This revised version was published online in July 2006 with corrections to the Cover Date.