Biochemical affinities between Cucumis hystrix Chakr. and two cultivated Cucumis species (C. sativus L. and C. melo L.) based on isozyme analysis

Isozymes of eight enzymes in Cucumis hystrix Chakr. and two cultivated Cucumis species (C. sativus L. and C. melo L.) were analysed electrophoretically to investigate the biosystematics of these three species. Cluster analysis using data from six enzymes indicated that considerable genetic distance existed between both C. hystrix and melon and between C. hystrix and cucumber. C. hystrix might be a key species for studying the evolution and taxonomy of genus Cucumis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Interspecific hybridization between Cicer arietinum and C. pinnatifidum

An interspecific hybrid between Cicer arietinum cv. GL 769 and a wild species C. pinnatifidum was obtained after emasculation, pollination and application of growth regulators. Ovules were cultured and embryos were later dissected to obtain hybrid plants. These plants were albinos and morphologically resembled C. pinnatifidum. Shrivelled seeds were also obtained in 2% of the crosses, which on germination gave rise to albino plants. These plants did not survive beyond 20 days. The hybrid nature of these plants was confirmed by esterase isozyme studies. Hybrid shoots obtained from germinating embryos were cultured on modified ML-6 medium with BAP 2 mg/1, IAA 0.5 mg/1, where they turned green after 3–4 weeks. Transmission electron microscopy (TEM) studies on leaf sections from green hybrid shoots showed an improvement in the chloroplast structure, with better organized grana.     

Effects of Temperature on Pollen Tube Growth and Fruit Set in Reciprocal Crosses Between Cucumis sativus and C. metuliferus

Interspecific hybridization between Cucumis sativus and C. metuliferus may be used to introduce disease resistance into the cultivated cucumber. Hybrid plant development, however, is restricted by strong barriers to crossing. The present study was undertaken to examine temperature effects on pollen tube growth and fruit set in reciprocal crosses between C. sativus and C. metuliferus. When crossing C. sativus×C. metuliferus, at 20°C, pollen tube growth was arrested in the stylar region of the pistil and no fruits containing embryos were found. However, at 23° and 26°C, C. metuliferus pollen tubes penetrated into the C. sativus ovules. The enhanced pollen tube penetration was correlated with improved embryo development. Although pre-fertilization barriers were overcome, post-fertilization barriers caused abortion of hybrid embryo development at the globular stage. No effects of higher temperatures were found in the cross C. metuliferus×C. sativus; Pollen tubes were generally arrested in the stylar region of the pistil.     

Meiotic chromosome behaviour in an allotriploid derived from an amphidiploid × diploid mating in Cucumis

A synthetic amphidiploid species Cucumis hyriviis Chen & Kirkbride (2n = 4x = 38; genome designated as HHCC) has recently been created from an interspecific mating between C. sativus L. (2n = 2x = 14: genome designated as CC) and C. hystrx. Chakr. (2n = 2x = 24; genome designated as HH). This amphidiploid is resistant to root knot nematode, tolerant to low irradiance, and has higher nutritional value than standard processing cucumber cultivars. An allotriploid (2n = 3x = 26; HCC) was derived from a cross between C. sativus L. and C. hytivus Chen & Kirkbride. Diploid meiotic behaviour in C. sativus and C. hystrix involves the development of seven and 12 metaphase bivalents respectively. In the derived allotriploid. univalents. bivalents, and trivalents (at relatively low frequency) were observed at metaphase I indicating that some homeologues from the C and H genomes can synapse. Based on a comparative karyotype analysis of cucumber (i.e. chromosome size and pairing behaviour) and aliotriploid plants, the seven bivalents observed at metaphase I were ascertained to be cucumber homologues, while the 12 univalents were of C. hystrix origin thus confirming the allotriploid karyotypic constitution to be HCC. On average, the frequency of trivalents was 0.24 at diakinesis and 0.22 in 100 meiocytes at metaphase I. indicating the possibility of genetic exchange due to the homoeology between the C and H genomes. After simultaneous cytokinesis, only polyads were observed in pollen mother cells (PMCs) at telophase II, which led to the production of sterile pollen grains. Multi‐polarization of chromosomes was dominant at anaphase II. However. in about 20% PMCs. chromosomes separated to form a 7C + 1‐2H complement, suggesting a possible method for the production of alien addition cucumber‐C hystrix lines through further backcrossing of the allotriploid to diploid cucumber.     

分葱对黄瓜、萝卜和白菜的化感作用

以黄瓜（Cucumis sativus L.）,萝卜（Raphanus sativus L.）和白菜（Brassica chinensis L.）3种蔬菜作物为受体,通过种子萌发试验及幼苗生长试验,对分葱（Allium fistulosum L.var.caespitosum Makino）根系及其地上部水浸液的化感作用进行了初步研究。结果表明：分葱根系和地上部水浸液对黄瓜、萝卜和白菜具有一定的化感作用。对黄瓜和萝卜的萌发有一定的抑制作用,而对其幼苗生长有一定的促进作用;对白菜的萌发表现为低浓度促进高浓度抑制,而对其幼苗生长有一定的抑制作用。因此,在蔬菜栽培制度中,分葱可与黄瓜和萝卜进行合理的轮作与间套作;但可能不适宜与白菜进行轮作或间套作。     

Interspecific hybridization between Vigna unguiculata (L.) Walp. and V. vexillata (L.) A. Rich. through in vitro embryo culture

Immature embryos resulting from the cross V. vexillata × V. unguiculata were cultured on MS medium supplemented with 2,4-D (2 mg/l) and resulted in embryogenic calli. Thirteen hybrid regenerants were obtained via organogenesis by subculturing the calli on MS medium supplemented with BAP (2 mg/l) + adenine sulphate (40 mg/l) + CH (500 mg/l) + cowpea tender pod extract (10%). The interspecific regenerants showed intermediate morphological traits between the parents for leaf shape, pod colour and seed coat colour. The hybrid plants inherited stem, leaf and pod hairiness of the wild species which could serve as a mechanical barrier against viral vectors. Electrophoretic studies of two isozyme systems, peroxidase and esterase, also confirms the hybrid nature of the regenerants as they expressed unique bands of both parents. Cytological study of the meiotic chromosomes revealed high frequency of univalent formation in the hybrids suggesting that the genomes of the parental species are structurally differentiated. The hybrid regenerants exhibited high enzyme activity for three enzymes viz., peroxidase, polyphenol oxidase and phenyl alanine ammonia lyase over the cultivated parent which may be useful in conferring resistance against viral pathogens. This revised version was published online in July 2006 with corrections to the Cover Date.     

Efficient hybridization between Lycopersicon esculentum and L. peruvianum via 'embryo rescue' and in vitro propagation

For the transfer of valuable traits from wild species into the cultivated tomato, excised globular-stage embryos 13 and 15 days after pollination (DAP) were cultured in vitro. Plants were regenerated from interspecific crosses of Lycopersicon esculentum cv. ‘Early pink’×L. peruvianum PI270435, and backcrosses of L. esculentum‘Giban (JF) No. 1’× (‘Early pink’× PI270435). Somatic embryos and single cotyledons emerged on hypocotyl sections of the embryos. Five to nine plantlets per embryo were obtained by clonal propagation. The hybrid nature of the plants is confirmed by comparing hybrids and parents in their ability to regenerate shoots from leaf segments in vitro, by comparing plant morphology and characteristics and by chromosome number. This study describes an efficient ‘embryo rescue’ method, as well as somatic embryogenesis by clonal propagation. A novel and simple method for the characterization of the interspecific hybrids is also reported.     

The effect of ovule age on ovary-slice culture and ovule culture in intraspecific and interspecific crosses with Tulipa gesneriana L.

The efficiency of two embryo rescue techniques, direct ovule culture and ovary-slice culture followed by ovule culture, is studied in crosses with Tulipa gesneriana as maternal genotype. The germination percentages increased, in most cases, significantly with increasing ages of te ovary-slices and ovules at the start of the cultures. The low number of embryos recovered at early culture dates is caused by a higher rate of embryo abortion and by retarded embryo development. The germination percentages for ovary-slice culture followed by ovule culture was mostly comparable to direct ovule culture. Unique hybrids have been obtained from the crosses T. gesneriana × T. agenensis and T. gesneriana × T. praestans. This revised version was published online in July 2006 with corrections to the Cover Date.     

黄瓜离体器官再生体系的优化

优化通过器官直接再生方式的黄瓜离体再生体系,为遗传转化奠定基础。以‘长春密刺’黄瓜的子叶节为外植体,探讨在黄瓜再生过程中,适宜的无菌苗获得培养基、适宜的外植体类型、无菌苗的适宜苗态、不定芽诱导培养基和芽伸长培养基中适宜的激素组合与比例。结果表明,最适的无菌苗获得培养基为1/2MS+30 g/L蔗糖+7 g/L琼脂(pH=5.8);不同外植体的再生率为子叶节>下胚轴>子叶;子叶完全出壳但未展平的无菌苗比其他苗态的再生率高,子叶展平后,再生率迅速下降;当6-BA（6-苄基氨基嘌呤）和ABA（脱落酸）浓度一定时,最适的AgNO₃为2 mg/L;当AgNO₃浓度一定时,6-BA和ABA浓度的最佳组合为1.5 mg/L 6-BA+0.5 mg/L ABA;不定芽诱导的最适培养基为MS+1.5 mg/L 6-BA+0.5 mg/L ABA+2 mg/L AgNO₃,出芽率为90%,每外植体出芽数为3.5;芽伸长培养基中加入0.10 mg/L 6-BA,能够促进再生芽的伸长。本研究成功优化了黄瓜的再生体系,得到了健壮的黄瓜成株。     

Combining ability between lines of Cucumis sativus L. and Cucumis sativus var. hardwickii (R.) Alef

Summary Three Cucumis sativus L. (CS) lines adapted to USA growing conditions were used as female parents in crosses to one line each of seven Cucumis sativus var. hardwickii (R.) Alef. (CH) accessions used as male parents to determine the relative combining ability of the CH accessions for six horticultural characters. The 21 F₁ progenies were grown in a randomized complete block design in two locations and evaluated for fruit number, lateral branch number, fruit length, fruit length/diameter ratio, number of female flowering nodes, and days to anthesis. General combining ability (GCA) was significant for all characters in each location. Specific combining ability (SCA) was significant for all characters except days to anthesis in one location and lateral branch number in both locations. Significant location × combining ability interactions were also evident for several characters. Results suggest that CH may be useful for improving fruit yield in commercial cucumber, but that limitations may lie in the attainment of acceptable fruit.Research supported by the College of Agric. and Life Sci., Univ. of Wisconsin, Madison; USDA, ARS, and by funds from Pickle Packers Intl., Inc. Mention of a trademark, proprietary product, or vendor does not constitute a guarantee of warranty of the product by the USDA and does not imply its improval to the exclusion of the other products or vendors that may also be suitable.     

Incompatibility in diploid and tetraploid crosses of Cucumis sativus and Cucumis metuliferus

The African horned cucumber (Cucumis metuliferus Naud.; 2x = 2n = 24) contains genes that can confer resistance to many important cucumber (C. sativus L.; 2x = 2n = 14) pests [e.g., root-knotnematode, Meloidogyne incognita (Kofoid & White) Chitwood]. Cucumber is highly susceptible to this root-knot nematode species, and a recent screening of C. sativus accessions in the U.S. National Plant Germplasm collection did not identify sources of resistance. Thus,autotetraploids of Cucumis sativus and C. metuliferus were created to recover fertile resistant interspecific progeny. Autotetraploids were obtained at the highest rate when seeds were immersed in 0.5% colchicine for a period of 6 to 8 hrs. Treatment durations less than 6 hrs produced few tetraploids, and durations of 10 hrs or more were lethal to seeds or developing seedlings. Crosses between C. sativus and C. metuliferus were made using diploid and tetraploid lines in all possible combinations, including reciprocals. Fruit development occurred in crosses when diploid and tetraploid C. sativus were used as the female parent. However, seeds developed only in fruit of C. sativus (4n) ×C. metuliferus (2n) crossings. Seeds from these crosses, however,were flat and not viable. No fruit development occurred in crosses whereC. metuliferus was used as the female parent. This revised version was published online in August 2006 with corrections to the Cover Date.     

Ovule and embryo culture to obtain hybrids from interspecific incompatible pollinations in chickpea

Many of the wild species of chickpea were not accessible to the improvement of chickpea due to cross incompatibility. In these interspecific incompatible crosses, fertilization takes place but the embryo aborts a few days later. The only way to obtain hybrid plants is by the application of growth regulators to pollinated pistils to prevent initial pod abscission and to save the aborting hybrid embryos by embryo rescue techniques. Although there are a few papers on regeneration from different explants of chickpea, information on embryo rescue techniques is not available. The paper summarises the embryo rescue techniques developed for chickpea, by the use of which hybrid plants between C. arietinum and C. pinnatifidum were produced. The paper also emphasises the effect of genotype to successfully obtain hybrids. The morphology of the hybrid plants resembled the male parent in leaf structure and growth habit. The colour of the flowers produced on the hybrid plant was pale violet, resembling the male parent whose flowers were violet in colour. The flower colour of the female parent was white. This revised version was published online in August 2006 with corrections to the Cover Date.     

Development and characterization of interspecific hybrids of Trifolium alexandrinum×T. apertum using embryo rescue

This is the first report on the development of interspecific hybrids between Trifolium alexandrinum and T. apertum using embryo rescue and characterization of F₁ plants. T. apertum was used as the male parent and T. alexandrinum as the female parent. Development of interspecific hybrids under natural conditions is not successful and so embryo rescue was attempted. Of the several combinations tried, pollination 2 days after emasculation and embryos rescued 11 days after pollination was found to be the best. For embryo culture, EC3 medium consisting of MS basal supplemented with 2.3 μM kinetin and 3% sucrose was used. Germinated embryos were transferred to LSP3 medium 25 days after inoculation wherein most of the cultures showed multiple shoots that were split and subcultured on RL1 medium for rooting. After hardening, about 75% of hybrids were successfully transferred to the field. The hybrids, in general, showed morphological traits intermediate between the two parents; however, a few hybrids showed better growth than either parent. Some F₁ plants were almost 3 weeks later in flowering than the female parent. Pollen fertility among these plants ranged from 78 to approximately 100%. Chromosomal associations at diakinesis and isozyme banding patterns for acid phosphatase (ACP), superoxide dismutase (SOD) and peroxidase also confirmed the hybrid nature of the plants.     

Interspecific hybridization between sunflower and wild perennial Helianthus species via embryo rescue

Interspecific crosses were made between the cultivated sunflower inbred line HA89 and accessions of five wild perennial Helianthus species (H. giganteus L., H. laevigatus T. & G., H. resinosus Small, H. pauciflorus Nutt. and H. decapetalus L.) resistant to broomrape ( Orobanche cernua Loefl.). Using the genetic male-sterile isogenic version of that line as female, successful reciprocal crosses were also obtained with all these wild species except with H. decapetalus. Five-day-old hybrid embryos were excised and cultured in vitro. In all cases, few mature plants were obtained from embryos in early developmental stages (early heart and globular) but up to 28% mature plants were obtained from embryos in later stages of development. Very immature embryos were difficult to excise without damage. Hybrid embryos and mature plants were obtained from all five wild species. Interspecific hybrid embryos from different wild species showed distinct developmental potentials, the proportion of hybrid embryos in different developmental stages varying among species. Differences in the proportion of hybrid embryos at the several developmental stages were also obtained for the reciprocal crosses (cultivated × wild), which showed higher proportion of fully developed embryos. Hybrids involving H. giganteus and cultivated sunflower were difficult to obtain without the use of embryo culture. Embryo culture proved to be an useful tool to overcome post-zygotic hybrid incompatibility in different Helianthus spp., and facilitated interspecific transfer of resistance to O. cernua. This revised version was published online in July 2006 with corrections to the Cover Date.     

The Gene Pools of the Grasspea (Lathyrus sativus L.)

Interspecific hybridization between the grasspea, Lathyrus sativus, and 15 wild species in Section Lathyrus is reported. Only two species, L. amphicarpos and L. cicera produced viable F¹ hybrids with low fertility when crossed with L. sativus as male parent. Crosses with six other species produced pods following pollination when L. sativus was the male, but seedlings were inviable, seeds did not germinate or pods were empty or had totally shrivelled seeds. When L. sativus was the female parent, only one cross with L. gorgoni produced an F¹, but the seedling was inviable. The germplasm resources of the grasspea are identified, with L. amphicarpos and L. cicera placed in the secondary gene pool and the other species in the tertiary gene pool. The definition of these germplasm resources is discussed in terms of grasspea improvement through plant breeding.     

黄瓜作图亲本间分子标记的多态性分析

利用AFLP,SRAP和SSR3种不同类型的分子标记技术,研究了作图亲本F-3与HZL04-1间的多态性。结果表明,这3种标记技术多态性检测效率不同,其中,AFLP揭示作图亲本间多态性效率最高,SRAP次之,SSR多态性检测效率最低。选择高效揭示多态性的分子标记技术是提高黄瓜遗传图谱构建效率的关键。分析探讨了几种分子标记技术的优缺点,认为应用AFLP,SRAP构建黄瓜遗传图谱为最佳,SSR可以作为补充的标记。     

Cytoplasmic male sterility in Nicotiana tabacum L. obtained through interspecific hybridization

Three new sources of cytoplasmic male sterility in tobacco with interspecific origin are reported. The wild species Nicotiana excelcior Black (n = 19), N. amplexicaulis Burbidge (n = 18) and N. rustica L. (n = 24) are used as donor of cytoplasm. Nuclear donor is N. tabacum L. (n = 24). The genetic effects of cytoplasmic-nuclear interactions of the studied species are discussed. Our experiments prove that N. excelcior, N. amplexicaulis and N. rustica are carriers of cytoplasmic genes for male sterility that express in combination with nuclear genes of N. tabacum. This revised version was published online in July 2006 with corrections to the Cover Date.     

Main Factors Affecting Cucumber (Cucumis sativus L.) Haploid Embryo Development and Haploid Plant Characteristics

This report gives the first detailed analysis of cucumber haploid plants and the first evaluation of genotype and season effects on haploid embryo development. Female flowers of four F1 varieties and two inbred lines were pollinated with 300 Gy-treated pollen and three-week old embryos were cultured in vitro on E20A medium. The variety ‘Polan’ produced the greatest number of embryos (1.34 per 100 seeds), 51 % of which were capable of further development. The number of embryos able to develop was four times higher in summer than in spring. All the plants obtained, except one, were haploids or aneuhaploids. Haploid plants grew slowly and were sterile, although they occasionally set seedless fruits. The length of the stems, leaves and guard cells was smaller than those of diploids.     

Analysis of reproductive isolation between pearl millet (Pennisetum glaucum (L.) R.Br.) and P. ramosum, P. schweinfurthii, P. squamulatum, Cenchrus ciliaris

Crosses between pearl millet lines and Pennisetum ramosum, P. schweinfurthii, P. squamulatum or Cenchrus ciliaris were observed for the frequency and development of zygotes, the possibility of embryo rescue, and the fertility of F1 hybrids obtained. Eight per cent of the ovules from diploid millet × P. ramosum crosses showed small embryos which could not be rescued. However, 59% of the ovules from tetraploid millet × P. ramosum crosses showed well-developed embryos that were easy to rescue 14 days after pollination. F1 hybrids were male sterile but female fertile when pollinated by diploid millet. Both diploid and tetraploid millet ovules showed the presence of hybrid zygotes after pollination with P. schweinfurthii at rates ranging from 25% to 45%. The diploid millet× P. schweinfurthii hybrid zygotes often developed almost normal seeds giving, without embryo rescue, totally sterile plants. The tetraploid millet × P. schweinfurthii hybrid embryos were normal but the endosperm was severely defective. A hybrid obtained by embryo rescue was totally sterile. A diploid millet-P. schweinfurthii amphidiploid was obtained by somatic embryogenesis associated with colchicine treatment during callogenesis. This amphiploid plant was male sterile, but gave many seeds when pollinated by a tetraploid millet and few seeds when pollinated by a diploid millet. P. squamulatum pollinating diploid millets produced proembryos with large undifferentiated endosperms in 73% of the ovules. A normal seed set was observed on tetraploid millets pollinated by P. squamulatum and the resulting F1 hybrids were partially male and female fertile. Backcrosses of these hybrids were much more fertile when pollination was from a tetraploid millet rather than from a diploid millet. C. ciliaris pollinating a diploid millet showed, in 60% of the ovules, proembryos and endosperms similar to those observed with P. squamulatum and no hybrid could be rescued. Crosses with a tetraploid millet could not be attempted due to the pistil-pollen incompatibility of tetraploid millets available with C. ciliaris. Ploidy levels of mating partners do not seem to influence pistil-pollen compatibility, but play a major role in post-zygotic abortion. With adequate ploidy levels of parents, and embryo rescue, it seems that the pearl millet gene pool can be considerably enlarged by germplasm from many other species.     

Investigations into the Barrier(s) to Interspecific Hybridization between Cicer arietinum L. and eight other Annual Cicer species

Barrier(s) to interspecific hybridization between the cultivated chickpea, Cicer arietinum L., and eight other annual wild species, i.e. C. reticulatum Lad., C. echinospermum Dav., C. pinnatifidum J. and S., C. judaicum Boiss., C. bijugum Rech., C. chorassanicum (Bge) M. Pop., C. yamashitae Kit. and C. cuneatum Rich., were investigated. In general, good pollen germination and pollen tube growth were observed in all eight crosses and their reciprocals. En spite of a few pollen cube growth abnormalities in most crosses, pollen tube penetration into the ovule and, thus, fertilization was observed in all cross combinations. However, differences were observed in the time from pollination to fertilization, not only between different interspecific crosses but also between reciprocals of a particular interspecific cross. The crossability barrier is, therefore, believed to be due to factor(s) operating after fertilization.