Breaking the intergeneric crossing barrier in papaya using sucrose treatment

A breeding programme was undertaken using Carica papaya var. Surya and Vasconcellea cauliflora with a view to raise progenies resistant to ‘papaya ringspot virus’ (PRSV). Earlier studies have clearly demonstrated the cross incompatibility between these two genera. Hence, an attempt was made to break this barrier using sucrose. The pollen of V. cauliflora was collected and pollination was carried out by smearing sucrose solution in the concentrations of 1, 2, 3, 4 and 5% on to the stigmatic surface of the flower. At 5% sucrose concentration, maximum viable seed set (13.73) was obtained. Sucrose at 5% was observed to break the intergeneric barrier by enhancing the pollen germination. There was drastic reduction in the effect of sucrose with the decrease in the concentration levels. The pollen germination studies carried out with and without sucrose clearly demonstrated the efficacy of sucrose in enhancing pollen germination and pollen tube growth. The intervarietal hybridization carried out between the varieties Surya and Pusa Dwarf showed 91.7% set of the fruits with 300 viable seeds per fruit. The hybridity of the progenies was confirmed using ISSR primers by the amplification of DNA from progenies and parents. Four primers UBC 807, 810, 814 and 861 clearly amplified male specific bands, which were present in progenies, but absent in female parent.     

Hybridization among Epimedium (Berberidaceae) species native to China

The hybridization of fourteen plant populations belonging to seven Epimedium species native to China was studied by self-cross, infra-population cross, inter-population cross within species, and interspecific cross. Self-pollination studies on nine populations indicated high incompatibility; capsule-set rates were higher than zero in only three (4.61–6.76%). Interspecific cross-pollinations demonstrated high crossabilities in most cross combination (15.38–92.44% capsule-set rate), and the F₁ seeds possessed high germination rates (>20%). The F₁ hybrids of three interspecific cross combinations were raised to maturity. The morphology, karyomorphology of somatic cells, and pollen mother cell (PMC) meiosis of these F₁ plants revealed that they were all highly fertile (>76.10% in pollen viability), that there were few structural differences in chromosomes among species, and that most PMCs had 6 bivalents at MI. Abnormal chromosomal behaviors occurred in a minority, including chromosome bridges, unequal segregation of chromosome number, lagging chromosomes, and micronuclei. A series of experimental crosses provided strong evidence for an outbreeding system and a weak internal barrier to hybridizations in the taxa studied.     

Intergeneric hybridizations between Opisthopappus taihangensis and Chrysanthemum lavandulifolium

For the first time, reciprocal intergeneric hybridizations were produced between Opisthopappus taihangensis and Chrysanthemum lavandulifolium without emasculation. About 20% seed set was similarly obtained from reciprocal hybridization. Only 5 and 6 of the viable plants observed from 45 and 78 seedling survived present some conspicuous intermediate characteristics. Phenotypic evaluation among the progenies of the parents and the putative hybrids was performed carefully since an average of 5.3% seed set was produced in the type of self-pollination using pollen from the same flower and >10% seed set was similarly obtained in the types of self-pollination using pollen from different flowers in a plant and flowers in individual plants from different seeds. One individual of each hybrid shared the inflorescence habit with the pollen plant was confirmed further by the sequence of ncpGS. The two hybrids might be used as bridges of breeding of multi-generic hybrids.     

Genetic transformation of Prunus domestica L. using the hpt gene coding for hygromycin resistance as the selectable marker

The study and development of transformation technology with new selection schemes is important for various fundamental studies and for crop trait improvement via genetic engineering. Here we have shown that hygromycin resistance is an effective system for plum genetic transformation. Embryonic axes of mature seeds were co-cultivated with Agrobacterium strain LBA4404 containing the pC1381 plasmid carrying the hygromycin phosphotranferase gene (hpt) and β-glucuronidase (GUS) gene or with strain EHA105 containing the plasmid pC1301 carrying the same marker and reporter genes. The latter strain containing a pC2301 plasmid carrying the neomycin phosphotransferase gene (nptII) gene was used as a control. Infected explants were placed on shoot induction medium containing either 5 mg L⁻¹ hygromycin or 75 mg L⁻¹ kanamycin for selection. Green shoots developed from the explants under hygromycin pressure. These shoots showed continued and vigorous growth and development upon transfer onto fresh hygromycin medium. PCR using hpt sequence primers, and Southern blot analysis using a probe from the hpt gene, confirmed the presence of the transgenes and their stable integration in regenerated plants. Full transgenic plants were obtained in a greenhouse. Hygromycin selection was very effective and no escapes were observed. The study demonstrated that hygromycin resistance can be used as an effective selectable marker for plum transformation. The new system developed here is important and useful for multiple gene transformation in plum.     

Increased regeneration capacity in spinach lines obtained by in vitro self-fertilisation

Somatic embryos (SEs) were induced from apical sections of the lateral roots of spinach seedlings (1 cm), which were cultivated on solid Murashige and Skoog (MS) medium with 20 μM α-naphthaleneacetic acid and 5 μM gibberellic acid. Apical shoots of the same lines were isolated and cultivated on plant growth regulator-free medium. The regeneration capacities of seedlings randomly chosen from a population were quite low and variable, and only 4 out of 30 lines responded at the frequency of 85–100%, with 6.96–9.96 SEs per explant and up to 347 SEs per seedling over a 12-week period. These SEs were isolated and maintained on medium with 5 μM kinetin. Plants derived from seedlings’ apical shoot and SEs self-fertilised in vitro and set seeds, and these seedlings (S₁) were used to induce regeneration. Similarly, S₂–S₄ seedlings were obtained and the regeneration capacities of 23 S₁, 23 S₂, 17 S₃ and 5 S₄-seedlings were compared to parental lines. Of these, four S₃ and S₄ lines with extremely high regeneration capacities were selected. These lines exhibited 78–139 fold higher embryo-forming capacities than the mother plant, and produced 38.9–68.3 SEs per explant and 1339–2181 SEs per seedling during the same time period. In addition, the process of somatic embryogenesis began 2–4 weeks earlier in these lines, and root explants taken from SE-derived plants of these lines retained high and stable regeneration capacities, and therefore may be ideal material for genetic transformation.     

Interspecific variations in seed germination of Corylopsis

This study was initiated to investigate the differences in germination percentages and rates between Corylopsis coreana Uyeki and Corylopsis sinensis var. calvescens Rehder & E.H. Wilson following a warm stratification (WS) and cold stratification (CS), and to study the effect of different WS temperatures interacting with different durations of CS. Warm stratification at 10 °C, 15 °C, 20 °C, and 25 °C was given for 1 month (1 M 10 °C, 15 °C, 20 °C, and 25 °C WS) followed by 0 M, 1 M, 2 M, and 3 M of CS at 5 °C (0 M, 1 M, 2 M, 3 M CS) and seeds were germinated in an air conditioned greenhouse maintained at 18.5 °C/18 °C. On average, less than 1% of C. coreana seeds germinated when sown without any WS and CS or with 1 M 15 °C, 20 °C, and 25 °C WS without CS treatment. However, 26% C. coreana seeds germinated after 1 M 10 °C WS without any CS treatment. Germination was not affected by WS temperatures when followed by 2 M 5 °C CS. It is concluded that C. coreana exhibited low seed germination at 10 °C and that this temperature could be considered the upper limit of CS for C. coreana. Only 2 M CS was required for more than 90% seeds to germinate. However, C. sinensis var. calvescens required longer than 3 M CS for more than 29% seeds to germinate. This clearly shows that there is an interspecific variation in optimum dormancy-breaking requirements.     

A strain of Bacillus subtilis stimulates sunflower growth (Helianthus annuus L.) temporarily

Preliminary studies showed that a Bacillus subtilis strain stimulates plant growth. We investigated how inoculating seeds of a sunflower cultivar (Helianthus annuus L.) with this strain stimulated plant growth, soil properties and emissions of greenhouse gasses, i.e. carbon dioxide (CO₂) and nitrous oxide (N₂O), when cultivated in a greenhouse. Unfertilized sunflowers or fertilized with urea served as controls. After one month, root length and fresh and dry root weight of the sunflower was significantly higher in the bacteria amended plant than in the urea and unfertilized plants. However, at harvest, no positive effect was observed. The number of seeds per plant and seed weight was not significantly different between the treatments, but total plant N was significantly higher in urea-amended plants than in unfertilized plants. The CO₂ production rate was not affected by treatment, but the N₂O emission rate was significantly higher in soil amended with urea plus bacteria soil compared to the unfertilized treatments. It was found that the B. subtilis strain used in this study had a positive, but only temporarily effect on growth of the sunflower cultivar used.     

Towards crop improvement in bell pepper (Capsicum annuum L.): Transgenics (uid A::hpt II) by a tissue-culture-independent Agrobacterium-mediated in planta approach

Agrobacterium tumefaciens-mediated transformation has been one of the methods used to generate transgenic plants in bell pepper. An alternate transformation method that avoids/minimizes tissue culture would be beneficial for the improvement of bell pepper due to its recalcitrant nature. In this report, transgenic bell pepper plants have been developed by a tissue-culture-independent A. tumefaciens-mediated in planta transformation procedure. In the present study, two open pollinated varieties viz., Arka Gaurav and Arka Mohini were used for transformation. Agrobacterium strain EHA105 harboring the binary vector pCAMBIA1301 that carries the genes for β-glucuronidase (uid A) and hygromycin phosphotransferase II (hpt II) was used for transformation. GUS histochemical analysis of T₀ and T₁ plants at various stages of growth followed by molecular analysis using PCR, Southern analysis and RT-PCR allowed selection of transgenics. The method resulted in 17.8% and 11.4% of the T₀ plants in Arka Gaurav and Arka Mohini being selected as chimeric and 35.0% and 29.7%, respectively, were identified as stable transformants in the T₁ generation based on PCR analysis.     

Midday depression of photosynthesis is related with carboxylation efficiency decrease and D1 degradation in bayberry (Myrica rubra) plants

Diurnal patterns of photosynthesis in two-year-old Myrica rubra young trees under natural conditions were studied by measuring gas exchange, chlorophyll fluorescence parameters and D1 protein. When measured on a clear day, the diurnal changes of net photosynthetic rate (P_n), stomatal conductance (G_s) and apparent quantum yield (AQY) show two daily maxima; the maximum value occurred at about 09:00 h, then declined, and reached the lowest values at about 13:00 h then increased to reach the second maxima at about 15:00 h. However, with the consistent decline of P_n and G_s in the afternoon, the ratio of intercellular CO₂ concentration (C_i) to atmospheric CO₂ concentration (C_a) increased. In addition, carboxylation efficiency (CE) and RUBP regeneration declined as the afternoon progressed. In the morning, the maximum yield of fluorescence after dark adaptation (F_m) and maximal photochemical efficiency of PSII (F_v/F_m) decreased continuously until it reached its minimum at 13:00 h, while the reverse occurred in the later afternoon. The quantum yield of PSII (Φ_PSII) declined after 09:00 h; in contrast, initial fluorescence (F_o) increased. A decrease in the rate of Q_A reduction and an increase in inactive PSII reaction centers was observed as the day progressed. Non-photochemical quenching (qN) and its slow-relaxing (qS) component increased at about midday, while the fast-relaxing component (qF) declined. The amount of inactive PSII centers was significantly enhanced, while F_v/F_m, Φ_PSII, qS and rate of Q_A reduction were significantly reduced by DTT (dithiothreitol), an inhibitor of the xanthophyll cycle. The D1 protein was significantly degraded at 13:00 h relative to that at 09:00 h during the course of the day. These results suggest that stomatal and non-stomatal limitations, which decreased carboxylation and photochemical efficiency, may cause the midday depression; and that non-stomatal limitations may be due to the decrease in RuBPCase activity and degradation of D1 protein, which causes decreased photoprotection at midday.     

Germination of Passiflora mollissima (Kunth) L.H.Bailey, Passiflora tricuspis Mast. and Passiflora nov sp. seeds

Passiflora mollissima, Passiflora tricuspis and Passiflora nov sp. are three passion fruit species occurring in Bolivia. Germination percentages and rates were determined for 11 different treatments. Per species, germination of 100 seeds was monitored every 3 days, during 90 days. Germination started after 9 days and 50% of final germination was reached within a month or less. Successful, recommended methods for P. mollissima are removing the basal point of seeds (27% germination) or removing the basal point in combination with pre-soaking seeds for 48 h in 50 ppm GA₃ (18%). Pre-soaking seeds for 24 h in 400 ppm GA₃ (42%) and removal of the basal point in combination with pre-soaking seeds for 48 h in 50 ppm GA₃ (36%) are suggested methods to improve germination of P. nov sp. Removing the apical point of P. tricuspis seeds resulted in maximal germination (57%). No unique treatment gave satisfactory results for the three species tested. Exogenous dormancy, probably a combination of mechanical and chemical dormancy is present in the three species studied. Presence of physical dormancy was found in P. mollissima.     

Mycorrhizal colonization improves onion (Allium cepa L.) yield and water use efficiency under water deficit condition

Most plants benefit from mycorrhizal symbiosis through improvement of water status and nutrient uptake. A factorial experiment with complete randomized blocks design was carried out in greenhouse at Tabriz University, Iran in 2005–2006. Experimental treatments were (a) irrigation interval (7, 9 and 11 days), (b) soil condition (sterile and non-sterile) and (c) arbuscular mycorrhizal fungi (AMF) species (Glomus versiforme, Glomus intraradices, Glomus etunicatum) and non-mycorrhizal (NM) plants as control. Onion (Allium cepa L. cv. Azar-shahr) seeds were sown in sterile nursery and inoculated with fungi species. One nursery left uninoculated as control. Nine weeks old seedlings then were transplanted to the pots. Average pre-irrigation soil water contents reached to about 67, 61.6 and 57.5% of FC corresponding to 7, 9 and 11 days irrigation intervals, respectively. At onion bulb maturity stage (192 days after transplanting), yield, water use efficiency (WUE) and yield response factor (K_y) were determined. The results indicated that AMF colonization increased soil water depletion significantly. G. versiforme under both soil conditions (sterile and non-sterile) and G. etunicatum in sterile soil depleted soil water effectively (P < 0.05). Mycorrhizal fungi improved WUE significantly (P < 0.0001) in both soil conditions. It raised by G. versiforme about 2.4-fold (0.289 g mm⁻¹) in comparison with the control (0.117 g mm⁻¹). G. intraradices and G. etunicatum also had significantly higher WUE than control. Apparently water deficit in 11-day irrigation interval led to lower yield and WUE compared to 9-day interval; the later resulted highest WUE (0.254 g mm⁻¹). Mycorrhizal plants increased seasonal ET significantly due to enhancing in plant growth; G. versiforme in both sterile and non-sterile soil and G. etunicatum in sterile soil had the highest ET. Bulb yield was influenced by irrigation period and fungi species. G. versiforme produced higher yield than other treatments (135.27 g/pot). Mycorrhizal plants in 11-day irrigation interval in spite of suffering from water stress had more bulb yield than non-mycorrhizal plants in all irrigation intervals. Yield in general was higher in 9-day treatments than other irrigating internals significantly (P < 0.05). Onion yield response factor (K_y) was decreased by AMF colonization; implying that symbiosed plants become less responsive to water deficit (longer irrigation interval) compared to the control ones.     

Development of seedless and Mal Secco tolerant mutant lemons through budwood irradiation

Mal secco (caused by Phoma tracheiphila (Petri) Kantsch. and Gik.) is the most destructive fungal disease of lemon plantations worldwide and seedless lemons would be preferred by most consumers. Five dosage levels, 0, 3, 5, 7, and 9 kiloradian (krad), of cobalt (⁶⁰Co) gamma irradiation were applied to budstick of ‘Kutdiken’ lemon (Citrus limon (L.) Burm. f.) clone KT-2A. Mutations were stabilized in three vegetative generations. Three hundred fifty-eight and 478 M1V3 (mutation one and vegetation three) plants were evaluated for seed number and mal secco tolerance in the field and the greenhouse, respectively. LD₅₀ was around 5 krad gamma irradiation for ‘Kutdiken’ lemon. The seed number varied from 0 to 34 per fruit. The level of mal secco tolerance also varied significantly among the plants from 1.0 (no symptom) to 4.3 (high level of disease occurance). The stable seedless and mal secco tolerant plants were obtained from 5 and 7 krad irradiation: the three mutants from 5 krad irradiation gave more lemon-like fruits, while 7 krad irradiation caused altered tree morphology and early maturation of fruits. This study shows considerable potential for lemon cultivar improvement aiming to obtain seedless and mal secco tolerant lemons.     

Seed inoculation with Azospirillum mitigates NaCl effects on lettuce

Data on the growth-promoting effects of Azospirillum on lettuce exposed to either normal or saline conditions, is scarce. Lactuca sativa L., cv Mantecosa seeds were colonized with A. brasilense Sp245 cells during imbibition. Germination percentages were determined after 7 d treatments with 0, 30, 50 or 80 mol m⁻³ NaCl. In another experiment, seeds germinated in Hoagland were irrigated for 30 d with 0, 30, 50 or 80 mol m⁻³ NaCl supplemented media. Vegetative growth proceeded in a growth chamber with a 13–11 h day–night cycle. Buffer-imbibed seeds were considered non-inoculated controls. Plant samples were taken at 0, 14, 20, and 30 d after the onset of NaCl treatments and dissected in aerial and root portions. The weights of both tissues were measured. Azospirillum-inoculated seeds had significantly higher germination percentages than controls in all treatments. Inoculated dried seeds stored up to 30 d maintained such characteristic in most of the treatments, particularly at 80 mol m⁻³ NaCl. Plants grown from inoculated seeds and irrigated with saline media displayed higher total fresh and dry weights and biomass partition to the aerial portion, than non-inoculated controls. Azospirillum-inoculated lettuce seeds had better germination and vegetative growth than non-inoculated controls after being exposed to NaCl.     

与黄瓜矮生基因连锁的ISSR标记及其SCAR转换

以黄瓜矮生品系D8与蔓生品系JIN5杂交并自交得到的124株F₂代为试材,应用ISSR分子标记技术和BSA（混合群体分组分析法）法寻找与黄瓜矮生基因连锁的分子标记。从80条ISSR引物中筛选到一个黄瓜矮生性状特异的多态性引物UBC818,经F₂代单株验证,UBC818的扩增片段在蔓生黄瓜JIN5植株中稳定出现,而在矮生黄瓜D8植株中没有。连锁关系分析表明,标记与黄瓜矮生基因间的遗传距离为11.1 cM。根据对该片段的序列分析结果重新设计了1对引物,将ISSR标记成功转化成了SCAR标记, 并命名为UBC818-S。     

The responses of photosynthetic capacity,chlorophyll fluorescence and chlorophyll content of nectarine (Prunus persica var. Nectarina Maxim) to greenhouse and field grown conditions

Three nectarine (Prunus persica var. Nectarina Maxim.) cultivars grown under solar-heated greenhouse and open-field in northwest China, were tested to evaluate their photosynthetic and chlorophyll fluorescence response to both growth conditions, and whether nectarine plants acclimate to the solar-heated greenhouse growth condition. Comparisons of light-saturated photosynthetic capacity (Amax) and CO₂-saturated photosynthetic capacity (RuBPmax) indicated that each cultivar (Z, Zao-Hongzhu; H, Hua-Guang; Y, Yan-Guang) maintained similar rates of light-saturated and CO₂-saturated carbon assimilation when grown in both conditions. The curve of diurnal variation of net photosynthetic (P_N) rate showed double peaks in open-field but single when grown in greenhouse. Compared with open-field-grown plants, a significant increase of daily average P_N was found in Z but decreased in Y in greenhouse. The diurnal variation of F_v/F_m indicate that plants grown in greenhouse experience less photoinhibition than in open-field condition. A reduction in chlorophyll (chl) a/b ratio in leaves of greenhouse grown plants with significant increase in chlorophyll (chl) b content were obtained. The results suggest that some nectarine cultivars have the ability to acclimate to the solar-heated greenhouse growth condition.     

Regeneration and transformation system in Mirabilis jalapa

Protocols for in vitro regeneration and production of in vitro-propagated plants and a transformation system were developed for Mirabilis jalapa (Nyctaginaceae). Among the types of explants and the different media tested, consistent shoot regeneration was obtained only from nodal segments grown in a regeneration medium consisting of Murshashige and Skoog medium supplemented with 2 mg l⁻¹ 6-benzyladenine, 2 mg l⁻¹ zeatin and 1 mg l⁻¹ indole acetic acid. Regeneration efficiency was dependent on the type of plant – white or pink flowers – used as the source of explants. Stable transformation was obtained following inoculation of nodal segments with Agrobacterium tumefasciens strain EHA105, which harbours the binary plasmid pAD1339 containing both nptII and gus genes under the control of the 35S promoter. Transformation was confirmed by PCR and Southern blot analysis of genomic DNA from mature regenerated plants. β-Glucuronidase (GUS) activity was observed only in tissues regenerated from in vitro-grown plants and not in tissues originating from greenhouse-grown plants. GUS expression was not uniform in regenerated leaves and showed a chimera pattern.     

In situ induction of chromosome doubling in vine cacti (Cactaceae)

Vine cacti have economic potential as exotic fruits in semi-arid and arid lands due to their high water use efficiency. The goal of this study was to attain autopolyploid plants by applying the antimitotic agents colchicine and oryzalin on axillary vegetative buds and germinating seeds. The diploid Hylocereus monacanthus and the tetraploid H. megalanthus; the interspecific triploid hybrid S-75, were studied. The effects of different concentrations and exposure times on bud survival and germination rate were recorded. A negative effect on vegetative bud survival was observed in the triploid hybrid. An inhibitory effect on germinating seeds was species specific, being greater on H. monacanthus with oryzalin and on H. megalanthus with colchicine. Putative polyploids were identified by comparing stomatal density between donor plants and plants arising from treated buds or seeds. Flow cytometric analysis and chromosome counts confirmed polyploidization in 14 autotetraploid H. monacanthus, 1 autohexaploid S-75 hybrid, and 14 autooctaploid H. megalanthus. This is the first report of artificial autopolyploidization in species of vine cactus, providing valuable novel plant material for further breeding programs.     

Development of ITS sequence based SCAR markers for discrimination of Paphiopedilum armeniacum, Paphiopedilum micranthum, Paphiopedilum delenatii and their hybrids

Paphiopedilum armeniacum, Paphiopedilum micranthum and Paphiopedilum delenatii are endangered orchid species. These three Paphiopedilum species and their hybrids are difficult to distinguish morphologically. In this study, rDNA-ITS (internal transcribed spacer) sequences were used to design species-specific SCAR (sequence characterized amplified regions) markers to distinguish P. armeniacum, P. micranthum, P. delenatii and their respective hybrids. The developed markers efficiently amplified 600 bp DNA product for P. armeniacum and its hybrids (SCAR-600armF/Pap-ITS2R), 300 bp product for P. delenatii and its hybrids (SCAR-300delF/Pap-ITS2R) and 700 bp product for P. micranthum and its hybrids (SCAR-700micF/Pap-ITS2R). The effectiveness of designed species-specific markers was also confirmed by using multiplex polymerase chain reaction amplification with a combination of developed three SCAR markers.     

Genetic structure of Buxus sinica var. parvifolia, a rare and endangered plant

Buxus sinica var. parvifolia, a rare and endangered tree species in some semitropics alpine areas of China, plays an important role in the maintenance of the landscape and ecosystem. In this study, RAPD and ISSR markers were used to investigate the genetic diversity and structure of five natural populations and one tamed population of B. sinica var. parvifolia. 21 RAPD primers amplified 209 bands with 167 (79.90%) polymorphic and 21 ISSR primers amplified 518 bands with 467 (90.15%) polymorphic. The genetic diversity, estimated by Shannon’ index, was 0.4343 (by RAPDs) and 0.3661 (by ISSRs). Both RAPD and ISSR analyses revealed a high level of genetic diversity in natural populations of B. sinica var. parvifolia. Furthermore, analysis of molecular variance (AMOVA) was used to apportion the variation within and between populations. The proportion of variation attributable to within-population differences was very high (69.2% by RAPDs; 84.51% by ISSRs). Moderate differentiation was detected among populations using RAPDs (30.80%), while only a small amount of variation (15.49%) was detected among populations using ISSRs. We suggest that the present genetic structure is due to high levels of environmental variability and gene flow, which still need further study to confirm. Conservation measures are suggested, including in situ and ex situ strategies, based on the observed population genetic information.     

Development of a male sterile eggplant by utilizing the cytoplasm of Solanum virginianum and a biparental transmission of chloroplast DNA in backcrossing

To develop a male sterile line of eggplant (Solanum melongena L.) utilizing the cytoplasm of S. virginianum, an interspecific F₁ hybrid between S. virginianum and S. melongena ‘Senryo Nigou’ was continuously backcrossed to S. melongena ‘Uttara’ using ‘Uttara’ as a recurrent pollen parent and four backcross generations, BC₁, BC₂, BC₃ and BC₄ were produced. All the plants in backcross generations were anther indehiscent although the F₁ hybrid, S. virginianum and S. melongena were dehiscent. Pollen stainability with acetic carmine and in vitro germination ability of pollen in all the backcross progenies were quite lower than those of the parental species. Fruit set percentage, number of seeds per fruit and seed germination rate were high in all the backcross progenies. The present results indicate that anther indehiscent type of functional male sterile line of eggplant could be developed by utilizing the cytoplasm of S. virginianum. PCR-RFLP analysis of chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) were performed to identify the organelle inheritance. The F₁ hybrid and all the backcross progenies displayed maternal inheritance of mtDNA. The cpDNA of the examined single BC₁ plant exhibited the recombinant cpDNA pattern of the parental F₁ hybrid and ‘Uttara’, indicating occurrence of the biparental inheritance of cpDNA. As all the BC₂, BC₃ and BC₄ progenies showed the same recombinant cpDNA patterns of the BC₁ plant, the recombinant cpDNA might be stable and harmonize with the nuclear genome of S. melongena. The present male sterile line can contribute to expand the male sterility source of eggplant.