• Title/Summary/Keyword: pollen culture

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Hybridization Using a New Male-sterile Germplasm as the Female Parent in Chinese Jujube

  • Wang, Jiu-rui;Cui, Xiu-mei;Dai, Li;Liu, Ping;Zhao, Jin;Liu, Meng-jun
    • Horticultural Science & Technology
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    • v.33 no.3
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    • pp.396-402
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    • 2015
  • This study identified a new male-sterile germplasm of Chinese jujube, named male-sterile No. 2 (JMS2), and achieved controlled hybridization using that germplasm as the female parent. The anthers of JMS2 before flower bud opening became shrunken, dingy yellow and much smaller than normal ones, and they changed to brown after anthesis. No pollen was observed in anthers of JMS2 and its male-sterile trait remained stable over different years. A total of 1,642 fruits were obtained from ten intra- and interspecific cross combinations via controlled hybridization from 2008 to 2012 using JMS2 as the female parent. Of those, 27.3% produced seeds, on average (0-72.6%). The rate of fruit with seed (RFS) was significantly different between cross combinations or years. Compared to other cross combinations, the RFS in the combination of JMS2 ${\times}$ 'Xingguang' (a Chinese jujube cultivar with high resistance to jujube witches' broom disease) and JMS2 ${\times}$ 'Xing16' (a sour jujube genotype) remained high in different years and reached means of 48.7 and 58.1%, respectively. Finally, 150 plantlets were regenerated from immature embryos, and 51 of them were randomly selected and identified to be authentic hybrids using amplified fragment length polymorphism (AFLP) markers. This is the first report of hybrids obtained from a cross between Chinese jujube and sour jujube.

Physio-chemical impacts on efficient germination in Platycodon grandiflorum for. duplex pollen culture

  • Kwon, Soo Jeong;Hwang, Ha Nule;Moon, Young Ja;Cho, Gab Yeon;Woo, Sun Hee;Boo, Hee Ock;Koo, Jin-Woog;Kim, Hag Hyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.170-170
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    • 2017
  • The highly valued ornamental plant, Platycodon grandiflorum for. duplex was generated by petaloidy of a calyx of Platycodon grandiflorum. The present investigation was executed to explore the several factors having effects on the germination of pollens with a view to acquire the underlying data for the artificial crossing to cultivate the species of Platycodon grandiflorum for. duplex. Both low and high temperature impaired the germination of Platycodon grandiflorum for. duplex pollens. The good germination rate was observed at the temperature of $25^{\circ}C$. The types and concentrations of carbon sources induced the differences in germination rate. The germination rate increased with the increasing concentration of sucrose and glucose except for fructose. Sucrose and glucose showed the highest results at the concentration of 20%. While fructose demonstrated the similar tendency to sucrose and glucose, it reduced the germination rate at the concentration of 20%. The highest germination rate was observed at the concentration of 15%. The appropriate carbon course for germination of pollens of Platycodon grandiflorum for. duplex was glucose of which germination rate was twice as high as that of sucrose and fructose. The germination rate was reduced substantially when the pH was close to alkali, and the potential germination rate was obtained at pH 6. Boric acid enhanced the germination rate at a lower concentration than the higher concentration.

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Effect of Embryo Morphology on Plant Development in Anther Cultures of Paeonia lactiflora Pall. (작약(Paeonia Lactiflora Pall.)의 약배양에서 형성된 배의 형태가 유식물 발달에 미치는 영향)

  • 손재근;권용삼;김경민
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.165-168
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    • 1995
  • The embryos formed from anther culture of peony exhibited divergent morphologies. The frequency of normal embryos with two cotyledons was about two times higher in the embryos formed through direct embryogenesis than those formed from callus. About 69% of the embryos with two cotyledons converted into normal plants, but convention rate of the abnormal embryos with one and three or four cotyledons was only 4 to 9%. The embryos of hem and bowling pin shape did not undergo development into normal plants.

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Consideration on the Scientific Analysis of Ancient Soil (고대 토양의 과학적 분석에 대한 고찰)

  • Seo, Min-Seok;Kim, Min-Hee;Chung, Yong-Jae
    • Korean Journal of Heritage: History & Science
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    • v.37
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    • pp.309-326
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    • 2004
  • There are some methods such as fatty acid analysis and microscope analysis of parasite egg and pollen and genetic analysis of ancient bacteria from ancient soil. The fatty acid analysis can examine whether some materials is human feces or animals. This is important thing to reconstruct ancient toilet culture pattern. The methods using TLC and GC-MS as organic chemistry is able to confirm ancient diet life style and nutritive conditions. The microscope analysis of ancient soil is able to confirm ancient parasite egg and pollen. It is possible to analogize ancient human diseases from this analysis. Also, genetic analysis is able to confirm genetic diversity and variation pattern of ancient organisms in archeological soil. Most of all, it is convinced of carrying through genetic preservation of exterminated ancient organisms. If archeological soils should be analysed through the natural scientific methods such as organic chemistry, soil science, microbiology, molecular biology, and genetics, this is helpful for us to understand and interpretation past historic event. And it is expected to perform an major role for understanding origin of ancient human and life style.

Studies on the Anther Culture of Some Woody Species (목본식물(木本植物)의 약배양(葯培養)에 관(關)한 연구(硏究))

  • Kim, Jai Saing
    • Journal of Korean Society of Forest Science
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    • v.13 no.1
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    • pp.25-39
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    • 1971
  • Recently successful induction of haploid plant by means of anther culture method has become a big topic among geneticists and plant breeders. The haploid plant can be used as a precious material for such basic researches as mutation or genetics. Once the haploid is obtained, production of homozygous plant is not a difficult problem. The method of producing homozygous plant can, also, be applied to the practical breeding works. When applied to the hybridization of self-fertilizing breeding period would be greatly shortened and in cross-fertilizing vegetables production of uniform hybrid seed would be very easily obtained. Last few years many scientists attempted anther cultures using various plant species, but it was successful only in several species. Unlike the other tissue cultures which use somatic organs or tissues as explants, anther culture seems to be very difficult because the plants or calli have to be induced from the haploid microspores or pollen grains. In the present experiment anther culture of fruit trees and ornamental shrubs of four genera and seven species was attemped. Anthers of Various stages ranging from tetrad and late microspore were cultured on the modified Murashige and Skoog's medium supplemented with various concentrations of auxins and kinetin as growth regulators. Handling of materials, sterilization, and other operations of culture were done by routine methods. The results were summarized as follows: 1. Calli were induced in the anthers of Forsythia Koreana Nak., Rhododendron mucronuratum Turcz., R. yedoense Max. var. Poukhanense Nak., and Prunus armeniaca L. var. ansu Max. No signs of callus were observed in Prunus persica Sieb. et Zucc. var. vurgaris Max., Pyrus ussuriensis var. macrostipes (Nak.), and Prunus salcina Lindley. 2. Calli were easily formed in any of the media with differing concentrations of auxins and kinetin. 3. In F. Koreana calli developed from anther surface and connective. Callus emerging out of anther locule was not observed. 4. Somatic calli arose from filament, connective, and inside of anther wall in R. mucronulatum. Many of the microspores accumulated starch grains. 5. The anther lobes located opposite the filament of R. yedoense turned easily to calli. This phenomenon was not observed in R. mucronulatum. Microspore embedded for a period in the medium became starch pollen. No callus was observed arising from microspore. 6. In P. armeniaca calli were not induced from somatic anther tissues. Instead, callus emerged out of anther locule rupturing the anther slit. Starch was not formed in the microspore. 7. In P. persica, Pyrus ussuriensis, and P. salcina, calli were not observed in the anthers examined more than 60 days after culture. Microspores of these species, however, were free of starch grains even after long period of subculture. 8. It was learned that somatic calli of the species examined arose usually from endothelium of anther wall, septum of two neighboring anther locules, parenchyma tissues of connectives, or anther lobes. 9. In the anther locule of P. armeniaca cultured long in medium, swollen microspores, polynucleate microspores, multicellular pollen grains, or callus mass were frequently observed, this indicating that the callus of this species was microspore-origin. 10. It was clarified that in P. armeniaca production of haploid plant by anther culture might be possible.

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Development of Pollen-Derived Embryos and Ploidy Level of Their Regenerated Plants in Paeonia ladctiflora Pall. (작약(Paeonia lactiflora Pall.) 화분에서 유래된 배의 발달과 분화식물체의 배수성)

  • 손재근;김광수;김경민
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.4
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    • pp.215-220
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    • 1994
  • Pollen-derived embryos cultured on the hormone-free medium showed a low germination frequency (12.5%) and poor growth response after germination. The greatest frequency of germination (81.3%) was obtained from the embryos cultured on medium with 0.3mg/L GA$_3$.The greatest frequency of generation (81.3) was obtained from embryos cultured on medium with 0.3mg/L GA$_3$. The embryos precultured for 20 days on medium with 0.3mg/L GA$_3$were transferred to the medium with various combination of hormones such as IAA, kinetin, zeatin, 6-benzylaminopurin (BA) and Gh$_3$. The germination frequency of cotyledonary stage embryos showed above 72% on media with all of the hormonal combinations, but the embryos germinated on medium with 2mg/L BA or 0.1mg/L kinetin and 0.3mg/L GA$_3$ developed more vigorously into plantlets than those of other hormonal combinations. Torpedo-stage embryos cultured on medium with 0.3 mg/L Gh$_3$ were pretreated for 8 weeks at 2-week intervals at 4$^{\circ}C$, The germination frequency of the cold-preheated embryos increased with the increment of pretreatment period from 2 to 8 weeks. The greatest frequency of germination (73.3%) was obtained from the embryos pretreated for 8 weeks at 4$^{\circ}C$. The chromosomes of the root-tip cells of W plane grown for 40 days after germination were observed. Most of the regenerated plants were haploid (55.8%) or diploid (315%), but triploid (1.3%), tetraploid (5.2%), or aneuploid (6.5%) were also detected among them.

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Interspecific Hybridization between Populus caspica L. × P. deltoids L 62/154 Using in vitro Embryo Development and Germination

  • Ali, Jafari mofidabadi;Mansooreh, Kamandloo;Hamid, Selamti
    • Journal of Forest and Environmental Science
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    • v.33 no.3
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    • pp.197-201
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    • 2017
  • Populus. caspica L. is an Iranian indigenous poplar species which naturally distributed in the northern part of country. Unfortunately, overuse has removed many of the stems of better form, so that natural stands now usually appear small and crook. Therefore genetic variation for selection of new superior clone of this species is needed. Conventional hybridization system is currently used to induce genetic variation in poplar species but incompatibility barriers have been observed between them. In vitro ovule embryo culture was used to overcome incompatibility obstacle for interspecific hybridization between Populus caspica L. with Populus deltoids L.62/75. Female flowers of Populus caspica L. have artificially been pollinated with pollen grain of P. deltoides 62/75 in one direction using twig and pot crossing system. Ovaries at different ages (7, 14 and 21 days after pollination) were disinfected through 70% ethanol for 1 minute, 5% of sodium-hypochlorite solution for fifteen min followed by three time rising with sterile distil-water. Isolated ovaries were then transferred to MS hormone free medium containing 30 and 60 g/L sucrose for embryo development and germination. Collected data have been analyzed by two factorial experimental designs. The results indicated that there were significant differences between age of embryos for development and germination at ${\alpha}=0.01%$. Highest embryo germination (45%) was observed from 21 days old ovaries. No significant differences were observed between MS culture media containing 30 and 60 g/L for percentages of ovary-embryo germination and number of germinated embryo per ovary at ${\alpha}=0.05%$. Fourteen percentage of embryo germination obtained in MS medium supplemented with 60 g/L sucrose, while only 35% of isolated ovaries were able to germinate in MS containing 30 g/L sucrose. Induced plantlets in 4 cm height were transferred into pots containing soilless (1:1:1 peat, per lit and vermiculite) medium for acclimatization. After successful acclimatization, plants were delivered to nursery.

Influences of Cold Pretreatment, Dark Condition, and Ficoll on the Callus Formation and Organ Differentiation in Anther Culture of Carnation (카네이션 약배양에 있어서 저온전처리, 암처리 및 Ficoll처리가 캘러스 형성 및 기관분화에 미치는 영향)

  • Lee, Su-Young;Kim, Jae-Yeong;Kim, Tae-Il;Ko, Jae-Young;Kim, Ki Sun
    • Horticultural Science & Technology
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    • v.17 no.2
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    • pp.138-140
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    • 1999
  • These studies were carried out to investigate the effects of cold pretreatment, dark condition, and Ficoll on the callus formation and organ differentiation in anther culture of carnation for the purpose of settling the techniques of anther culture of 'Rony' carnation. Diameter of flower buds containing anthers with microspores before or after uniuncleated stage was between 4 and 6 mm. A high percentage of callus formation and responding anthers were achieved by low temperature pretreatment at $4^{\circ}C$ for 7 days. Callus formation was promoted under dark condition. Roots were differentiated from calli formed under darkness. Ficoll didn't promote callus formation but promoted differentiation of shoots and roots.

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Hybridity Verification of Progenies Obtained from Ovule Culture by Using RAPD Markers in Reciprocal Crosses of Alstroemeria (알스트로메리아 배주배양을 통하여 획득한 정역교배 자손의 혼종성 분석)

  • Lee, Ja-Hyun;Joung, Youn-Hwa;Han, Tae-Ho
    • FLOWER RESEARCH JOURNAL
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    • v.19 no.4
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    • pp.231-237
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    • 2011
  • In this study, we performed ovule culture after reciprocal crosses of two Alstroemeria accessions and investigated genetic contribution of parents by using RAPD markers. The best method was half-ovule culture on MS medium supplemented with $60g{\cdot}L^{-1}$ sucrose and $2.2g{\cdot}L^{-1}$ gelrite at 14 days after pollination. Embryos began to germinate after 6 weeks of culture. The complete plantlets were formed after 4 months of culture. In eight progenies and two parental cultivars, 59 polymorphic bands were obtained out of 89 total bands by RAPD analysis using 7 primers. Eight $F_1$ progenies from the crosses between two accessions using reciprocal crosses showed 1:1 contribution of maternal and paternal parents. It is confirmed that $F_1$ progenies were obtained from parental accessions by using RAPD markers. We conclude this cross combination showed pre-fertilization barriers with incompatibility between stigma or style, and pollen because progeny number was different in each cross combination. Thereby, it warrants overcoming pre-fertilization barrier together with post-fertilization barrier in order to broaden the heterozygosity within progeny populations in Alstroemeria breeding program.

Flower and Microspore Development in 'Campbell Early' (Vitis labruscana) and 'Tamnara' (V. spp.) Grapes ('캠벨얼리'와 '탐나라' 포도의 꽃과 소포자 발달)

  • Yim, Bomi;Mun, Jeong-Hwan;Jeong, Young-Min;Hur, Youn Young;Yu, Hee-Ju
    • Horticultural Science & Technology
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    • v.33 no.3
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    • pp.420-428
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    • 2015
  • The majority of cultivated varieties of grape have perfect flowers that are clustered in an individual inflorescence. Grape flower has a single pistil, five stamens, a protective flower cap (calyptra), and a calyx. After fertilization, an individual flower develops into a single berry. Although there are a number of reported studies focusing on berry formation, berry enlargement, and sugar accumulation in grape, the morphological studies of flower, including gametophyte morphogenesis and structural change in floral organs, have not yet been studied in detail. In this study, we investigated the flower structure and development characteristics of grape using microscopy and defined the floral development stages 9 to 13 based on microspore or male gametophyte development stage from tetrad to mature pollen. We used seeded diploid table grapes 'Campbell Early' (Vitis labruscana) and 'Tamnara' (V. spp.) as plant materials. At floral development stage 9, pollen mother cells develop to tetrads. During floral development stages 10 to 11, unicellular microspore develop to mid bicellular pollen. At the end of floral stage 12, male gametophyte develops to mature tricelluar pollen. In floral stage 13, the flower cap falls off and flower bud opens. During floral development stages 9 to 12, there were no major changes in calyx length, whereas the length of the flower cap continuously increased. The flower cap-to-calyx length ratio was 2.0, 3.0, 4.5, and 6.5 at floral stages 9, 10, 11, and 12, respectively. The flower cap-to-calyx length ratio was consistent in the two grape cultivars, suggesting that the ratio is a morphological character representing floral development stage. This study provides a reference for determining floral development stage of the two grape cultivars. It will be useful for the determination of optimum time for microspore culture needed to generate doubled haploid lines and appropriate gibberellic acid treatment needed to induce parthenocarpic fruit development in 'Tamnara' grape.