Production and genetic characterization of interspecific hybrids among Crambe abyssinica, Crambe hispanica and Crambe kralikii
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In this paper, interspecific crosses among Crambe abyssinica, Crambe hispanica, and Crambe kralikii were reported.In the C. hispanica x C. abyssinica (H x A) cross, 118 F 1 hybrids were produced without embryo rescue, while 5 F 1 hybrids were obtained with embryo rescue, when C. hispanica was used as the female parent.In the reciprocal cross (A x H), 232 hybrids were obtained without embryo rescue.From more than 1000 C. kralikii flowers pollinated with pollen grains of C. abyssinica (K x A), only 2 F 1 hybrids were obtained with embryo rescue, whereas the reciprocal cross produced no hybrids, even with embryo rescue.The hybrids were confirmed at the morphological, cytological, and molecular levels.In the combinations of A x H and H x A, many BC 1 hybrids were obtained without embryo rescue.In contrast, in the K x A cross, only 7 BC 1 plants were obtained with embryo rescue, while no seed set was achieved under self-pollination or in backcrosses without embryo rescue.In the H x A F 1 hybrids, the pollen stainability was 65.4-86.0%,with an average of 76.9%.In comparison, the pollen viability of hybrids in the reciprocal cross (A x H) ranged from 66.2 to ©FUNPEC-RP www.funpecrp.com.brGenetics and Molecular Research 13 (3): 6995-7005 (2014) 81.1%, with an average of 75.4%.Fertile pollen grains were not found in the K x A F 1 hybrids.All F 1 hybrids of the 3 crosses (H x A, A x H, and K x A) had the expected 2n = 75 chromosomes.AFLP analyses indicated that all F 1 hybrids and their progenies had typical bands of the parents.These hybrids and progenies are anticipated to be valuable for future C. abyssinica improvement in breeding programs.Keywords:
Embryo rescue
Crambe
Reciprocal cross
Hand-pollination
Embryo rescue
Reciprocal cross
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SUMMARY Populations of Pontederia sagittata from the northern coastal plain of Mexico possess a tristylous breeding system. Styles and stamens are reciprocally arranged in the three floral morphs. Differences in anther size, pollen size and pollen production are associated with the three anther levels. Controlled pollinations performed in the glasshouse indicate that physiological self‐incompatibility accompanies floral trimorphism. Pollinations between morphs with anthers and stigmas at equivalent heights are highly productive of seed in comparison with self‐ or intra‐morph pollinations. In each floral morph, substantial differences in seed production result from self‐pollination with alternate anther levels. Overall, the mid‐styled morph exhibits weak self‐incompatibility whereas the long‐ and short‐styled morphs are more strongly self‐incompatible. In all pollen‐stigma combinations pollen germinated readily and pollen tubes penetrated stigmatic tissue. Inhibition of pollen tubes occurs in the style and possibly the ovary. The rate of pollen tube growth differs between anther levels following self‐pollination. The three floral morphs were represented in eight populations examined although there was a tendency for the long‐styled morph to be under‐represented. Strong pollen trimorphism allows the origin of pollen deposited on naturally pollinated stigmas to be determined. Considerable variation in the amount and composition of pollen deposited on stigmas by syrphid flies was recorded. The likelihood of legitimate pollination appears to be highest in the long‐styled morph, intermediate in the mid‐styled morph and lowest in the short‐styled morph. Tristyly in P. sagittata is compared with the closely related P. cor data and P. rotundifolia. Implications of weak self‐incompatibility in the mid‐styled morph for the breakdown of tristylous breeding systems in the Pontederiaceae are discussed.
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Premise of the Study Heterostyly, the reciprocal positioning of stigmas and anthers in different floral morphs, has long been thought to promote intermorph pollination. However, extensive intramorph pollination occurs commonly in heterostylous species, leading to recurrent questions about the functional and evolutionary significance of heterostyly. Methods To identify the sources of stigmatic pollen (autogamous [intraflower], geitonogamous [intraplant], vs. interplant), we emasculated either one flower or entire plants in experimental populations of the two closely related buckwheat species, distylous Fagopyrum esculentum and homostylous F. tataricum . Differences in pollen size allowed unambiguous identification of pollen on stigmas. Results Only 2.4% of F. tataricum pollen and 1.5% of F. esculentum pollen arrived successfully on compatible stigmas of other plants. In the former (homostylous) species, 71.3% of the pollen load on stigmas was autogamous, 10.8% was geitonogamous, and 17.9% was interplant. In the latter (distylous) species, 37.45% of the pollen on stigmas was autogamous, 13.8% was geitonogamous, 17.0% was intramorph, and 31.75% was intermorph. The amount of incompatible pollen arriving on stigmas was greatly decreased by both one‐flower and whole‐plant emasculations, and thus, the proportion of compatible pollen deposited increased with one‐flower emasculation and increased even more with whole‐plant emasculation. Conclusions Our quantification of pollen‐donor sources in these two species indicated that heterostyly in Fagopyrum esculentum provided a nearly 2‐fold fitness advantage (in terms of compatible pollination) over expected (random) pollen transfers between morphs. Because of reduced herkogamy, the homostylous F. tataricum was highly autogamous.
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Emasculation
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This research investigates the hybridization of species Lilium pumilum Delile, L. regale Wils., L. candidum L., L. monadelphum Bieb., L. martagon L., L. henryi Baker. and cultivars of Asiatic, Trumpet and Oriental hybrids. To overcome fertilization barriers in incongruous crosses, cut-style pollination and pollination by mixed pollen were performed. The hybrid plants were obtained by in vitro cultivation of excised immature embryos. Cut-style pollination was effi cient when Asiatic hybrids were pollinated by pollen of L. pumilum. Cut-style pollination and culture of excised embryo allowed receive few hybrids between Oriental hybrids and L. henryi or L. regale. Pollination of a female by a mixture of incompatible pollen was more effi cient than cut-style pollination. Hybrid plants with random paternity were produced using the pollen mixture.
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In this paper, interspecific crosses among Crambe abyssinica, Crambe hispanica, and Crambe kralikii were reported.In the C. hispanica x C. abyssinica (H x A) cross, 118 F 1 hybrids were produced without embryo rescue, while 5 F 1 hybrids were obtained with embryo rescue, when C. hispanica was used as the female parent.In the reciprocal cross (A x H), 232 hybrids were obtained without embryo rescue.From more than 1000 C. kralikii flowers pollinated with pollen grains of C. abyssinica (K x A), only 2 F 1 hybrids were obtained with embryo rescue, whereas the reciprocal cross produced no hybrids, even with embryo rescue.The hybrids were confirmed at the morphological, cytological, and molecular levels.In the combinations of A x H and H x A, many BC 1 hybrids were obtained without embryo rescue.In contrast, in the K x A cross, only 7 BC 1 plants were obtained with embryo rescue, while no seed set was achieved under self-pollination or in backcrosses without embryo rescue.In the H x A F 1 hybrids, the pollen stainability was 65.4-86.0%,with an average of 76.9%.In comparison, the pollen viability of hybrids in the reciprocal cross (A x H) ranged from 66.2 to ©FUNPEC-RP www.funpecrp.com.brGenetics and Molecular Research 13 (3): 6995-7005 (2014) 81.1%, with an average of 75.4%.Fertile pollen grains were not found in the K x A F 1 hybrids.All F 1 hybrids of the 3 crosses (H x A, A x H, and K x A) had the expected 2n = 75 chromosomes.AFLP analyses indicated that all F 1 hybrids and their progenies had typical bands of the parents.These hybrids and progenies are anticipated to be valuable for future C. abyssinica improvement in breeding programs.
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Teak flowers are weakly protandrous and pollen is shed within a few hours of flower opening. Pollen is tricolpate and 29 μm in diameter. The papillate stigma is of the wet type and is receptive from 1100–1300h. The style is hollow throughout its length. Nectar and pollen are the major floral rewards for pollinators. The major pollinators are Ceratina sp. which carry teak pollen on most parts of their bodies, especially the specialized hair structures (scopal brushes) on the tibia. The most effective pollination period in terms of flowers pollinated and pollen per flower is between 0900 and 1300h. At 1300h the number of pollen per flower is the highest, ranging from 1–36 (average 7). Pollen tubes grow very fast. Within 2 h after pollination 8% of the pollen tubes have reached the micropylar end of the ovule and pollen tubes first enter the embryo sac at 8 h. Only one to two pollen tubes enter the micropyles of a flower. Although 78% of flowers were pollinated in open-pollination, the low fruit set (3.5%) suggests that there are factors other than pollination limiting fruit set. The main factor appears to be a high amount of selfing, and self-incompatibility occurs when pollen tubes are arrested at the lower portion of the ovary.
Tectona
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Hybridization between the nearly extinct speciesDiplotaxis siettiana andBrassica juncea is prevented because of strong prefertilization barriers. Use of mentor pollen ofD. siettiana irradiated with 1000 Gy gamma radiation before the incompatible pollination led to fertilization. 5 d after pollination 17% ovules showed entry of pollen tubes, 10 d after pollination 27% ovules showed small globular embryos which grew no further. No embryos were found in control pollinations. Thus , use of irradiated mentor pollen brings about fertilization in this difficult cross and hybrids can be obtained if embryos are rescued.
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Reciprocal hybrids between an induced autotetraploid form of Glycine tomentosa (Benth.) (n=20) and G. tabacina (Labill) Benth. (n=40) were obtained. All F 1 hybrids were morphologically intermediate between the parents and were highly male and female sterile as estimated by pollen stainability and seed set. However, F 1 hybrids with G. tomentosa cytoplasm had lower pollen stainability and the anthers usually failed to dehisce. F 1 's with G. tabacina cytoplasm shed pollen abundantly. F 2 and backcross plants generally were lower in fertility than the F 1 's. In all comparisons made between G. tomentosa and G. tabacina cytoplasms the former was associated with lower fertility. Both species are members of the subgenus Leptocyamus and limited attempts to cross them with G. max (of the sub‐genus, Soja ) were unsuccessful. Cytological observations indicated three different genomes are involved in the two species crossed. G. tomentosa was designated 2(A 1 ) and G. tabacina 2(A 0 A 2 ). Genome A 1 is apparently more closely related to both genome A 0 and A 2 than A 0 and A 2 are related to each other.
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To introduce yellow, blue or orange pigmentation in the Rhododendron subgenus Tsutsusi (including R. simsii hybrids) by classical breeding, intersubgeneric pollination experiments involving Hymenanthes, Pentanthera and Rhododendron (including Vireya) species and hybrids were started. Prefertilization research by pollen tube staining revealed a different behaviour between bilateral crosses. Ovule culture was applied as a means to save a substantially larger amount of hybrid embryos from abortion. Though flower/seed capsule abortion eliminated many crosses in an early developmental stage, ovules of many crosses could be initiated in vitro on various time intervals after pollination. Germination was achieved on basal Rhododendron medium + 50 mg/l GA3. Green seedlings were obtained after Tsutsusi x Hymenanthes, Hymenanthes x Tsutsusi, Tsutsusi x Rhododendron, Rhododendron x Tsutsusi and Vireya x Tsutsusi pollination. Many seedlings have little growth vigour and next to green seedlings albino seedlings occur frequently, depending on the pollination type. The first true leaves of many assumed hybrids obviously lack chlorophyll. The ploidy level of Vireya x Tsutsusi and Tsutsusi x Pentanthera seedlings is intermediate, which strongly suggests their hybrid origin. Molecular analysis through SSR confirmed the interspecific charachter of seedlings in half of the crosses tested.
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Pollen tube growth rates in legitimate cross- and self-pollinations were compared by controlled hand pollination in two varieties of Pontederia cordata L. (Pontederiaceae). In each pollen–stigma combination pollen germinated readily on stigmas and pollen tubes penetrated stigmatic tissue. Pollen tube length in styles was positively correlated with pollen size. In most pollinations legitimate pollen tubes grew more rapidly in styles than self pollen tubes. This difference was statistically significant in most comparisons. Inhibition of incompatible pollen tubes was either in the style or ovary. It is suggested that the mechanisms of incompatibility may differ among the floral morphs and depend on the particular pollen size involved in pollinations. The observed differences in pollen tube growth of legitimate cross- and self-pollinations in P. cordata suggest that the species is primarily outbreeding.
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