Species hybridization in Trifolium

In this investigation, pollen tubes were traced in intraspecific and interspecific crosses in Trifolium. Pollen tube growth was slower in the interspecific crosses examined but no failure of pollen germination was observed. Incompatibility of the pollen tube and stylar tissue proved to be an important pre-fertilization barrier to compatibility and a parallel could be drawn between the success of the pollen and the success of vegetative grafting of the genotypes.     

Pollination and in vitro germination of seeds for interspecific hybridization of <Emphasis Type="Italic">Psidium guajava</Emphasis> and <Emphasis Type="Italic">Psidium cattleianum</Emphasis>

The genus Psidium includes important fruit crops. However, there are very few studies focusing on its reproductive biology, which limits the establishment of breeding programs. The present work investigated the reproductive biology of Psidium guajava and Psidium cattleianum in terms of compatibility of crossings between these two species aiming at interspecific hybridization because the latter species is an important source of resistance against the nematode Meloidogyne enterolobii. Several types of crosses were performed to understand the reproductive biology of these species, including the compatibility of intra- and interspecific crossings, using assisted in vivo germination of pollen grains on the stigma. In addition, the in vitro germination of both Psidium species was studied at different stages of fruit development to rescue young seeds to improve the chances of obtaining the hybrids. No fruits of 270 pollinations were obtained on guava buds at the pre-anthesis stage, regardless of the source of the pollen grain and the cultivar used as female genotype. Microscopic analyzes demonstrated the germination of pollen grains and pollen tube growth at crosses between guava cv. ‘Pedro Sato’ (P. guajava) and Psidium cattleianum. High germination percentages of Psidium cattleianum seeds were obtained in MS medium without sucrose or containing 15 g/L of this carbohydrate.     

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.     

Barriers to interspecific hybridization in Ulmus americana

Summary Crossability barriers between the tetraploid Ulmus americana and five other diploid elm species were investigated using both fluorescence and scanning electron microscopy. Both pollen germination and pollen tube development were inhibited in all interspecific crosses. Pollen tube behavior on the stigma of U. americana was unchanged when pollen of a colchicine-induced U. pumila tetraploid was substituted for diploid U. pumila pollen.Self-incompatibility was observed in one of two American elms studied and manifested itself via reduced pollen germination and inhibited pollen tube growth on the stigma surface.The results indicate that crossing barrier(s) between U. americana and other elm species occur at the stigma surface and operate independently of ploidy level. This conclusion is discussed in relation to current breeding efforts to transfer Dutch elm disease resistance into the American elm via interspecific hybridization with Eurasian elms.     

Investigation on pre-zygotic barriers in the interspecific crosses involving Gossypium barbadense and four diploid wild species

Interspecific hybridisation has played significant role in the improvement of economically useful traits such as productivity, earliness, fibre quality and resistance to pests and diseases in cotton. However, wide crosses are often limited by the operation of either pre or/and post-fertilization barriers. The present study on pollen tube behaviour of four diploid wild species viz., G. triphyllum, G. davidsonii, G. thurberi and G. armourianum in the pistils of G. barbadense was taken up to determine the extent of pre-zygotic barriers operating in these crosses. High rate of pollen germination (∼80%) and normal growth of pollen tubes were observed upon selfing of both G. barbadense and four wild species. Pollen tubes reached the pistils and fertilization was accomplished within 8 HAP in all the five parental species. In the interspecific crosses, pollen germination was normal in the cross involving G. armourianum and drastically inhibited in the crosses involving G. triphyllum, G. davidsonii and G. thurberi. Even though delayed pollen tube growth was a general feature in all the four crosses, successful fertilization was observed only in the cross involving G. armourianum. In crosses involving G. triphyllum, G. davidsonii and G. thurberi, pollen tubes failed to reach the ovary even at 24 HAP due to the presence of strong stylar and ovarian incompatibility. Abnormalities in pollen tubes such as swelling, twisting, intense callose plug formation, breakage, branching and reverse orientation were frequently observed among the incompatible crosses.     

Pollen germination and pollen tube growth following self-pollination and intra- and interspecific pollination of Medicago species

Summary Pollen germination and pollen tube growth after self- and intra- and interspecific pollinations of annual [M. scutellata (L.) Mill., M. disciformis DC., and M. rigidula (L.) All.] and perennial (M. sativa L.) Medicago species were studied using a Carl Zeiss microscope with a fluorescence filter attachment.Self-pollination of self-fertile annual and intraspecific pollination of cross-fertile perennials were characterized by high pollen germination and rapid normal pollen tube growth. In contrast pollen germination percentages were very low and many pollen tube growth abnormalities occurred following interspecific pollination and self-pollination of self-sterile plants. The time period from pollination to fertilization for interspecific crosses was about double that following self-pollination of the annual species. However, fertilization occurred frequently following interspecific pollinations. Much of the sterility was due to factor(s) operating after fertilization.Cooperative investigations of the Kansas Agricultural Experiment Station, Dept. of Agronomy and the U.S. Department of Agriculture, Agricultural Research Service. Contribution No. 82-582-J- Mention of a trademark name or proprietary product does not constitute a guarantee or warranty by the U.S. Department of Agriculture, and does not imply its approval over other products that also may be suitable.     

Increasing resistance to Fusarium crown and root rot in asparagus by gametophyte selection

In garden asparagus, Fusarium crown and root rot is the main cause of crop decline. Since chemical treatments are inefficient, efforts should focus on the development of resistant cultivars to control the disease. Toxic culture filtrate (TCF) of F. oxysporum has affected asparagus pollen germination and tube growth. Consequently, gametophyte selection was evaluated to ascertain if the application of selective agents at this level could increase selection efficiency. Two susceptible pistillate plants and one tolerant and one susceptible staminate plants were used in controlled crosses. Before pollination, a drop of a germination vehicle with TCF or without it was applied to the stigmas. Some pollinated pistils were fixed and analyzed by fluorescence microscopy; the rest were left on the plant for seed production. Fifty to 200 seeds were obtained per treatment combination (staminate plant x pistillate plant x pollination vehicle). The derived plantlets were inoculated in vitroand evaluated for disease symptoms. The application of TCF to stigmas reduced pollen germination and tube growth compared with untreated controls,regardless of the genotypic combination. Pollen germination and tube growth was poorer for the tolerant staminate genotype than for the susceptible one. When the TCF was applied, the number of seeds per pollination in comparison with the controls diminished only when the susceptible genotype was the pollinator. The percentage of affected root area of the progenies obtained after applying the TCF was lower than in the controls only when the tolerant genotype was the pollinator. Increasing Fusarium resistance in asparagus by means of gametophyte selection seems feasible. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effect of Abscisic Acid on Pollen Germination and Tube Growth of Maize Genotypes1

As a preliminary step to using gametophytic selection to obtain genotypes of maize, Zea mays L., differing for abscisic acid (ABA) content and/or sensitivity, the influence of ABA on the in vitro germination and tube growth of maize pollen and the presence of genetic variability for these traits was investigated. In a first experiment, pollen samples of three genotypes were germinated on substrates containing increasing amounts of ABA. Low ABA concentration (0.5 νM) significantly increased pollen germination and tube growth, while 500 νM ABA significantly decreased both traits, thus indicating that ABA can influence the growth of maize pollen in vitro. In a second experiment, eleven inbred lines were screened for the in vitro reactivity of their microgametophytes to ABA. The lines differed significantly for the response (% of control) of tube length, but not of pollen germination to ABA.     

In vitro embryo germination and interploidy hybridization of rose (Rosa sp)

In roses the problems associated with inter-specific breeding include low percent of seed set and lack or low percent of seed germination. Low seed set is usually due to non-amenable parents, which may have different ploidy level or other divergences that result in embryo abortion at early stages of development. Lack of seed germination is mostly attributed to the mechanical restrictions such as thick pericarp or the regulatory mechanisms such as the hormonal control of dormancy. The aims of the present investigation were to optimize in vitro embryo germination technique in rose and study the ploidy of progenies resulted from interploidy hybridizations. To optimize embryo germination, seeds were surface sterilized, whole pericarp and testa were removed and embryos were placed on half strength Murashige and Skoog media supplemented with different concentrations (0, 1, 2.5, 5 mg l^?1) of benzyladenine (BA) in combination with different concentrations (0, 0.5, 1, 1.5 mg l^?1) of gibberellic acid. The maximum percent of in vitro embryo germination (93.40 %) was observed on medium containing 2.5 mg l^?1 BA. In order to select the most fertile seed parents which could be used in interspecific hybridization, 11 commercial rose cultivars (R. × hybrida) were employed in 36 reciprocal crosses. Three rose cultivars including ‘Golden Celebration’, ‘Tess of the d’Urbervilles’ and ‘Molineux’ were selected as the maternal parents. The selected seed parents were employed in crosses with one rose species from Gallicanae section [R. damascena (2n = 4x)] and four rose species form Caninae section [R. orientalis (2n = 5x), R. iberica (2n = 5x) R. canina (2n = 5x) and R. pulverulenta (2n = 6x)]. The highest percent of hip set and in vitro embryo germination were observed in crosses between tetraploid rose cultivars and R. damascena. In all of the crosses with R. canina, the percent of hip and seed set was 0 %. However, in the crosses between tetraploid rose cultivars and other pentaploid or hexaploid rose species from Caninae section both triploid and tetraploid offsprings were attained. Future morphological analysis of the progenies is necessary to show to what extent progenies demonstrate the characteristics of the pollen parents from the Caninae section. Nevertheless, progenies from interploidy hybridizations would be beneficial in future breeding programs in order to expand the relatively small gene pool of roses.     

Compatibility relations between the edible carrot Daucus carota and D. pusillus, a related wild species from the Argentinian Pampas

To establish the feasibility of hybridization between the wild carrot species Daucus pusillus Michx. (2n = 2x = 22; 2n = 2x = 22 and 20), collected in the pampas grasslands of Argentina, and the edible carrot, Daucus carota L. (2n = 2x = 18), controlled pollinations were attempted on the plant. Due to the difficulties encountered, flowers of 12 accesions and three commercial cultivars were excised from individual plants and pollinated in Petri dishes following an incomplete diallel design. After processing, the pollinated pistils (four to six per genotypic combination) were observed under a microscope with UV light. Pollen tubes reaching the ovaries and/or the ovules (compatible relation) were observed in six out of nine D. pusillus × D. pusillus and seven out of 18 D. pusillus × D. carota genotypic combinations. In the eight D. carota × D. pusillus genotypic combinations, only ungerminated pollen, pollen not adhered to the stigmas or pollen tubes overlapping the stylar tissue were observed. Additional flowers were pollinated in a sample of compatible genotypic combinations and the pollinated pistils were in vitro culture to study embryo and endosperm development. Eight out of nine pistils from the intraspecific and nine out of 13 from the interspecific crosses enlarged to form apparently normal schizocarps. Histological analyses revealed normal development of embryo and endosperm. The breeding barriers between the two species are incomplete, making feasible the obtainment of interspecific hybrids by conventional techniques.     

Factors affecting seed set in the crosses between <Emphasis Type="Italic">Dendranthema grandiflorum</Emphasis> (Ramat.) Kitamura and its wild species

Reproductive barriers often exist in the crosses between Dendranthema grandiflorum (Ramat.) Kitamura and its wild species and seriously result in low seed set, consequently reducing breeding efficiency. For the purpose of revealing the factors leading to low seed set, we investigated pollen viability, germination behavior of pollen grains on stigmas and embryo development in the crosses between D. grandiflorum and three wild species, D. nankingense (Nakai) Tzvel., D. indicum (L.) Des Moul. and D. zawadskii (Herb.) Tzvel. using technique of paraffin section, and light, fluorescence and scanning electron microscopy. The results indicated pollen viability of three wild species ranged from 20 to 25%. In the cross between D. grandiflorum and D. nankingense, very few pollen grains germinated on stigmas after pollination and most of them germinated abnormally. In addition, normal embryos were observed in 12% ovaries at 8 days after pollination and thereafter all the embryos aborted. In other two crosses, many pollen grains germinated on stigmas and pollen tubes penetrated stigmas normally after pollination. Moreover, normal embryos were observed in over 50% ovaries from 8 to 15 days after pollination in the cross between D. grandiflorum and D. indicum, and seed set was 59%. In the cross between D. grandiflorum and D. zawadskii, normal embryos were observed in 52% ovaries at 8 days after pollination. After that, however, most embryos degenerated and seed set was only 9%. These data suggest that pollen viability has no significant effects on seed set of the three crosses. Very few germinated pollen grains on stigmas and abnormal growth of most pollen tubes before fertilization, and embryo abortion are the main factors causing failure of the cross between D. grandiflorum and D. nankingense, whereas only embryo abortion is a main factor resulting in low seed set in the cross between D. grandiflorum and D. zawadskii and no barriers occur in the cross between D. grandiflorum and D. indicum.     

Pollen–pistil interaction in the inter-specific crosses of Sesamum sp

The wild species of Sesamum have long been recognized as an important source of many valuable pest and disease resistance genes and other novel traits such as male sterility and plant type for the cultivated taxon. Utilization of these species in crop improvement is marginal because of their inherent low crossability with the cultigen. An understanding of the biological nature of incompatibility systems that impede hybridization offers tools for successful introgressions into cultivated sesame. The objective of this investigation was to observe pollen germination and pollen tube growth in cross pollinated S. indicum pistils as an indicator of pre-zygotic barriers operating in wide crosses involving S. indicum. Three wild species: S. alatum, S. radiatum and S. laciniatum were used as pollen donors in hybridization with S. indicum. In the cross S. indicum × S. alatum 85.23% of pollen grains germinated and pollen tubes reached ovaries within 8 h after pollination. In other two crosses, S. indicum × S. laciniatum and S. indicum × S. radiatum the level of incompatibility was high as evidenced by a reduced pollen germination and slow rate of pollen tube growth. Measures to circumvent incompatibility in these crosses to produce interspecific hybrids are discussed.     

苏丹草高粱及其杂种的细胞遗传学研究

苏丹草(S. sudanense)与高粱(S. bicolor)均为禾本科高粱属植物, 两者的杂种优势明显, 杂交种品质好, 抗逆性强, 在水产、畜禽养殖及资源利用与环境保护上有着广阔的开发利用前景, 但两者是否属于同一个种至今存在争议。本文采用去壁低渗-火焰干燥法, 分析了2份苏丹草、2份高粱及其3个杂种F₁有丝分裂核型, 观察了3个杂种F₁减数分裂染色体行为和2个杂种F₂体细胞染色体数目。结果表明, 苏丹草、高粱及其杂种F1均为1A核型, 但核型公式不完全相同, 苏丹草Sa为2n=18m+2sm(sat), 高粱3042A和3042A×Sa F₁为2n=20m, 其余材料均为2n=20m(sat)。苏丹草、高粱及其杂种F₁ 3者在10条染色体的绝对长臂、绝对短臂、绝对全长、臂比和相对全长上差异均不显著(P>0.05), 说明苏丹草与高粱在染色体长度上的变化不明显。杂种F₁花粉母细胞减数分裂, 终变期和中期I染色体核型和数目清晰可见(2n=2x=20), 配对行为规则; 棒状和环状二价体的频率因组合不同而异, Tx623A×S722 F₁、3042A×Sa F₁和Tx623A×Sa F₁棒状二价体频率分别为4.887、5.710和5.126, 环状二价体频率分别为5.113、 4.290和4.874; 在后期I, 配对的染色体能够正常分离。杂种F₂体细胞染色体数目为20(2n=20)。因此, 苏丹草与高粱的亲缘关系非常近。     

Prefertilization barriers to interspecific hybridization involving Gossypium hirsutum and four diploid wild species

Interspecific hybridization among species of cotton has lead to improvement in productivity, earliness, fibre quality and resistance to pests and diseases. However, wide crosses is often limited by the operation of either pre‐ or/and post‐fertilization barriers. An investigation on pollen tube behaviour of four wild species in the pistils of Gossypium hirsutum was taken up. Pollen germination was normal in crosses involving Gossypium triphyllum and Gossypium armourianum and markedly inhibited in the crosses involving Gossypium davidsonii and Gossypium thurberi. Pollen tubes reached the pistils and fertilization was accomplished within 8 h after pollination (HAP) in control cross. Even though delay in pollen tube was a common phenomenon in all the four crosses successful fertilization was observed in crosses involving G. triphyllum and G. armourianum, as they reached the ovary at 24 HAP. In crosses with G. davidsonii and G. thurberi, pollen tubes failed to reach the ovary even at 24 HAP indicating the presence of strong stylar and ovarian incompatibility. Measures to overcome such barriers to interspecific hybridization in the incompatible crosses are discussed.     

Pollen characteristics and stigma receptivity for <Emphasis Type="Italic">Anemone coronaria</Emphasis> L.

To obtain crossing products, an efficient pollination and subsequent fertilization is essential. This efficient pollination is achieved by pollen germination and tube growth. Here, these pollen characteristics of 2 genetically differentiating cultivars of Anemone coronaria L. were investigated in vitro. For the essential components boron, calcium and an osmoticant, only calcium showed to be crucial for pollen germination. Boron concentrations influenced the pollen tube length with a concentration of 100 mg l^?1 H₃BO₃ resulting in the longest pollen tubes. For the osmoticant sucrose, a concentration of 100 g l^?1 caused a significant positive effect on both pollen germination and pollen tube length for the 2 cultivars. The cultivars reacted similarly with respect to the investigated compounds. Next to this, the pollen development was correlated with 8 different flower stages in A. coronaria. As a final point, the germination of an optimized pollen germination medium was compared with in vivo pollen germination in cross-pollinations within the same cultivar (identified by aniline blue staining). For ‘Mistral Wine’, pollen germination percentage was lower in vitro than in vivo, while ‘Wicabri Blue’ pollen showed no significant difference in germination rates in vivo and in vitro. To achieve fertilization a following requisite is that the stigma is receptive. To study this, the most receptive female flower stage of the 8 different flower stages for A. coronaria was characterized by aniline blue staining. A. coronaria clearly showed protogyny.     

玉米与摩擦禾、薏苡的杂交不亲和性

采用荧光显微技术, 对摩擦禾、薏苡花粉在玉米柱头上的萌发和生长过程进行了观察。摩擦禾花粉粒在玉米柱头上均能萌发, 花粉管在柱头中伸长并到达花柱基部, 且可将雄配子送入胚囊内, 玉米果穗顶端有受精结实痕迹, 说明摩擦禾与玉米的杂交障碍不是杂交不亲和, 而是胚囊不亲和或杂种衰亡。薏苡花粉粒在玉米柱头也能萌发, 花粉管能伸入花柱, 但玉米与薏苡杂交生殖隔离较摩擦禾严格, 杂交极其困难, 杂交障碍为胚囊不亲和或花柱不亲和。玉米与薏苡杂交时, 薏米花粉管能到达玉米花柱基部, 而川谷花粉管却在花柱中停止生长, 杂交障碍与薏苡种类有关。玉米与薏苡杂交的花粉管异常率高于玉米与摩擦禾杂交花粉管异常率, 反映了玉米与摩擦禾的亲缘关系较与薏苡近。     

Failure in an interspecific cross between Solanum khasianum Clarke and Solanum mammosum L.

Summary Attempts to obtain a successful interspecific cross between Solanum khasianum Clarke and Solanum mammosum L. were unsuccessful, and the probable reasons for this failure were investigated. A reduced number of viable pollen grains and low receptivity of the stigmas of S. mammosum L. and heterostyly in both species are the probable reasons for the failure of the attempted interspecific crosses between S. khasianum Clarke and S. mammosum L.     

The role of pollen and stigma in determining the level of partial self-compatibility in brussels sprouts

Summary Brussels sprouts inbred plants, homozygous for the same recessive S-allele, were intercrossed in all combinations and the number of pollen tubes penetrating the stigmas of such crosses measured. The results of five such experiments showed that pollen as well as stigma is significant in determining the number of pollen tubes counted. It was also found that the capacity of a plant's stigmas to permit pollen tube penetration was independent of the capacity of its pollen to penetrate other plants' stigmas. Variation in pollen tube production between pollens of the plants tested was often greater than that between their stigmas.     

Elucidating the behaviour of pollen tubes in intra-and interspecific pollinations of Phaseolus vulgaris L. and P. coccineus Lam.

Summary The behaviour of pollen tubes in intra-and interspecific pollinations of P. vulgaris and P. coccineus has been examined using a combination of phase-contrast, and UV fluorescence microscopy. The results indicate that P. coccineus is self-compatible in terms of pollen tube growth, and that pollen tube inhibition is not responsible for the reciprocal differences in seed set which have been observed in crosses between these species.     

Marker-assisted breeding for rf1, a nuclear gene controlling A1 CMS in sorghum (Sorghum bicolor L. Moench)

A study on marker-assisted selection (MAS) for the rf1 gene, which controls pollen sterility in the sorghum A1 cytoplasm, was conducted on the offspring population of two crosses between a maintainer line, BTx-622, and two sweet sorghum lines, BJ-299 and Lunen-2, to test the effectiveness of the MAS method and develop maintainer lines with sweet and juicy stalks and corresponding cytoplasmic-nuclear male sterility (CMS) lines. The simple sequence repeat marker Xtxp18 exhibited a high accuracy (95.098 %) for selecting recessive homozygotes for the rf1 gene. The segregation ratio matched the expected ratio calculated according to the reported genetic distance in the F₂ population of the two crosses used. Finally, four excellent maintainer lines/CMS line pairs (F₅/BC₃) with high stalk juice and stalk juice sugar contents were developed. The MAS method based on Xtxp18 for the sorghum rf1 gene could be used for hybrid breeding programs at a low cost in the future.