scholarly journals Seed Development of Cypripedium debile Rchb. f. in Relation to Asymbiotic Germination

HortScience ◽  
2012 ◽  
Vol 47 (10) ◽  
pp. 1495-1498 ◽  
Author(s):  
Rebecca C.-C. Hsu ◽  
Yung-I Lee
Keyword(s):  
Culture Media ◽  
Nile Red ◽  
Basal Medium ◽  
Germination Percentage ◽  
Seed Collection ◽  
Cultural Conditions ◽  
Immature Seeds ◽  
Hydrophobic Nature ◽  
Slipper Orchid ◽  
Mature Seeds

The histological and histochemical changes in developing seeds of Cypripedium debile Rchb. f., a native slipper orchid species with horticultural potential, were investigated. The effects of timing for seed collection, culture media, and cultural conditions were also examined. The optimum germination percentage occurred when mature seeds were collected and sowed on 1/4 Murashige and Skoog basal medium. Besides, the liquid culture promoted germination of mature seeds. This finding is contrary to most other Cypripedium species, which are relatively easy to germinate with immature seeds. Moreover, two notable cytological changes of C. debile were observed. First, the suspensor cell protruded beyond the micropyle opening of the inner seedcoat, making the inner seedcoat not substantial. Second, Nile red staining indicated that the deposition of cuticular material on the seedcoat was fragmentary. It is proposed that the less hydrophobic nature of the seedcoat makes mature seeds of C. debile easier to obtain water and nutrients for germination.

scholarly journals Asymbiotic germination of Vanilla planifolia in relation to the timing of seed collection and seed pretreatments

2021 ◽  
Vol 62 (1) ◽  
Author(s):  
Chih-Hsin Yeh ◽  
Kai-Yi Chen ◽  
Yung-I. Lee
Keyword(s):  
Seed Germination ◽  
Seed Development ◽  
Seed Coat ◽  
Germination Percentage ◽  
Vanilla Planifolia ◽  
Outermost Layer ◽  
Seed Collection ◽  
Seed Pretreatment ◽  
Immature Seeds ◽  
Mature Seeds

Abstract Background Vanilla planifolia is an important tropical orchid for production of natural vanilla flavor. Traditionally, V. planifolia is propagated by stem cuttings, which produces identical genotype that are sensitive to virulent pathogens. However, propagation with seed germination of V. planifolia is intricate and unstable because the seed coat is extremely hard with strong hydrophobic nature. A better understanding of seed development, especially the formation of impermeable seed coat would provide insights into seed propagation and conservation of genetic resources of Vanilla. Results We found that soaking mature seeds in 4% sodium hypochlorite solution from 75 to 90 min significantly increased germination. For the culture of immature seeds, the seed collection at 45 days after pollination (DAP) had the highest germination percentage. We then investigated the anatomical features during seed development that associated with the effect of seed pretreatment on raising seed germination percentage. The 45-DAP immature seeds have developed globular embryos and the thickened non-lignified cell wall at the outermost layer of the outer seed coat. Seeds at 60 DAP and subsequent stages germinated poorly. As the seed approached maturity, the cell wall of the outermost layer of the outer seed coat became lignified and finally compressed into a thick envelope at maturity. On toluidine blue O staining, the wall of outer seed coat stained greenish blue, indicating the presence of phenolic compounds. As well, on Nile red staining, a cuticular substance was detected in the surface wall of the embryo proper and the innermost wall of the inner seed coat. Conclusion We report a reliable protocol for seed pretreatment of mature seeds and for immature seeds culture based on a defined time schedule of V. plantifolia seed development. The window for successful germination of culturing immature seed was short. The quick accumulation of lignin, phenolics and/or phytomelanins in the seed coat may seriously inhibit seed germination after 45 DAP. As seeds matured, the thickened and lignified seed coat formed an impermeable envelope surrounding the embryo, which may play an important role in inducing dormancy. Further studies covering different maturity of green capsules are required to understand the optimal seed maturity and germination of seeds.

scholarly journals Embryo Development of Cypripedium formosanum in Relation to Seed Germination In Vitro

2005 ◽  
Vol 130 (5) ◽  
pp. 747-753 ◽  
Author(s):  
Yung-I Lee ◽  
Nean Lee ◽  
Edward C. Yeung ◽  
Mei-Chu Chung
Keyword(s):  
Seed Germination ◽  
Embryo Development ◽  
Developmental Stages ◽  
Culture Media ◽  
Nile Red ◽  
Germination Percentage ◽  
Zygotic Embryogenesis ◽  
Seed Collection ◽  
Significant Difference

This investigation documents the key anatomical features in embryo development of Cypripedium formosanum Hayata, in association with the ability of embryos to germinate in vitro, and examines the effects of culture media and seed pretreatments on seed germination. A better understanding of zygotic embryogenesis for the Cypripedium L. species would provide insights into subsequent germination events and aid in the in vitro propagation of these endangered species. In seeds collected at 60 days after pollination (DAP), soon after fertilization, no germination was recorded. The best overall germination was found at 90 DAP (≈70%), at which time early globular to globular embryos with a single-celled suspensors can be observed. After 135 DAP, the seeds germinated poorly. At this time the inner integument shrinks and forms a tight layer, which encloses the embryo, the so-called “carapace.” Using Nile red stain, a cuticular substance was detected in the carapace, which may play a role in the impermeability of the mature seed and may help the seeds survive in the stringent environment. At maturity (after 210 DAP), the embryo proper has an average size of eight cells along its length and six cells across the width. Lipids and proteins are the main storage products within the embryo. To improve seed germination, experiments were conducted to test the suitability of various media and pretreatments of seeds. When different media were used, except for the Harvais medium at 120 DAP, there was no significant difference in seed germination at three different developmental stages tested. Soaking mature seeds in 1% NaOCl or treating them with ultrasound may slightly increase the germination percentage. For seed germination, our results indicate that the timing of seed collection outweighs the composition of medium and the seed pretreatments.

scholarly journals Asymbiotic Germination of Mature Seeds and the Seed Development of Vanilla Planifolia

2021 ◽  
Author(s):  
Chih-Hsin Yeh ◽  
Kai-Yi Chen ◽  
Yung-I Lee
Keyword(s):  
Cell Wall ◽  
Seed Germination ◽  
Seed Development ◽  
Seed Coat ◽  
Germination Percentage ◽  
Vanilla Planifolia ◽  
Outermost Layer ◽  
Seed Pretreatment ◽  
Immature Seeds ◽  
Mature Seeds

Abstract Background: Vanilla planifolia is an important tropical orchid for production of natural vanilla flavor. Traditionally, V. planifolia is propagated by stem cuttings, which produces identical genotype that are sensitive to virulent pathogens. However, sexual propagation with seed germination of V. planifolia is intricate and unstable because of the extremely hard seed coat. A better understanding of seed development, especially the formation of impermeable seed coat would provide insights into seed propagation and conservation of genetic resources of Vanilla.Results: We found that soaking mature seeds in 4 % sodium hypochlorite solution from 75 to 90 min significantly increased germination and that immature seeds collected at 45 days after pollination (DAP) had the highest germination percentage. We then investigated the anatomical features during seed development that associated with the effect of seed pretreatment on raising seed germination percentage. The 45-DAP immature seeds have developed globular embryos and the thickened non-lignified cell wall at the outermost layer of the outer seed coat. After 60 DAP, the cell wall of the outermost layer of the outer seed coat became lignified and finally compressed into a thick envelope. These features matches the significant decreases of immature seed germination percentage after 60 DAP. Conclusion: We report a reliable protocol for seed pretreatment of mature seeds and for immature seeds culture based on a defined time schedule of V. plantifolia seed development. The thickened and lignified seed coat formed an impermeable envelope surrounding the embryo, and might play an important role in seed dormancy of V. plantifolia.

scholarly journals Effects of Desiccation and Storage Temperature on Seed Germination in Kapok

2016 ◽  
Vol 26 (1) ◽  
pp. 83-88
Author(s):  
YanLing Zheng ◽  
GaoJuan Zhao ◽  
HuanCheng Ma
Keyword(s):  
Seed Germination ◽  
Storage Temperature ◽  
Seed Viability ◽  
Germination Percentage ◽  
Seed Vigor ◽  
Deciduous Tree ◽  
Seed Moisture Content ◽  
Seed Collection ◽  
And Storage ◽  
Mature Seeds

Kapok (Bombax ceiba) is a deciduous tree that can grow in the dry-hot valley of southwestern China where its natural regeneration by seedlings is difficult. As mature fruit split open and seeds disperse by wind, it is difficult to collect fully mature seeds. The effects of seed moisture content (MC) and storage temperatures on seed germination of dark-brown seeds collected from split fruit and light-brown seeds collected ≈10–15 days earlier than the time of fruit split were studied to determine the effective germplasm preservation via the seeds. Dark-brown mature seeds could tolerate desiccation to less than 5% MC and could tolerate −20 and −80 °C. Seeds of kapok showed orthodox storage behavior. They can be stored at subzero temperatures with low MC for a long time. For light-brown seeds, germination percentage (GP), germination index (GI), seedling fresh weight (SFW), and vigor index (VI) decreased significantly after seed desiccation. Germination percentage of light-brown seeds with different MC increased to a different extent after being stored at different temperatures for 1 year (76% to 99%), compared with the fresh seeds (73%). Storing fresh seeds at 4 °C was most favorable to keep seed viability and seed vigor of light-brown seeds. Seed collection could be done several days earlier than the time of fruit burst to ensure increased quantity of collected seeds.

scholarly journals Analysis of Polyhydroxyalkanoates Granules in Haloferax mediterranei by Double-Fluorescence Staining with Nile Red and SYBR Green by Confocal Fluorescence Microscopy

Polymers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1582
Author(s):  
Verónica Cánovas ◽  
Salvador Garcia-Chumillas ◽  
Fuensanta Monzó ◽  
Lorena Simó-Cabrera ◽  
Carmen Fernández-Ayuso ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Culture Media ◽  
Nile Red ◽  
Sybr Green ◽  
Confocal Fluorescence Microscopy ◽  
Fluorescence Staining ◽  
Haloferax Mediterranei ◽  
Optimized Protocol ◽  
Confocal Fluorescence ◽  
Pha Production

Haloferaxmediterranei is a haloarchaeon of high interest in biotechnology because it produces and mobilizes intracellular polyhydroxyalkanoate (PHA) granules during growth under stress conditions (limitation of phosphorous in the culture media), among other interesting metabolites (enzymes, carotenoids, etc.). The capability of PHA production by microbes can be monitored with the use of staining-based methods. However, the staining of haloarchaea cells is a challenging task; firstly, due to the high ionic strength of the medium, which is inappropriate for most of dyes, and secondly, due to the low permeability of the haloarchaea S-layer to macromolecules. In this work, Haloferax mediterranei is used as a halophilic archaeon model to describe an optimized protocol for the visualization and analysis of intracellular PHA granules in living cells. The method is based on double-fluorescence staining using Nile red and SYBR Green by confocal fluorescence microscopy. Thanks to this method, the capability of PHA production by new haloarchaea isolates could be easily monitored.

scholarly journals Ethylene and Lipoxygenase In Relation to Afterripening of Dormant NC-13 Peanut Seeds1,2

1985 ◽  
Vol 12 (1) ◽  
pp. 45-49 ◽  
Author(s):  
D. L. Ketring ◽  
H. E. Pattee
Keyword(s):  
Heat Treatment ◽  
Cold Storage ◽  
Ethylene Production ◽  
Sharp Increase ◽  
Gradual Increase ◽  
The Other ◽  
Immature Seeds ◽  
Positive Correlations ◽  
Mature Seeds

Abstract Following harvest, peanuts are usually subjected to a period of storage. During storage biochemical changes are known to occur. The objectives of this study were to determine the changes and relationship in ethylene production, germination, and lipoxygenase (LG) activity during cold storage of dormant NC-13 peanut seeds. Two seed lots (SL) were used: one grown in Oklahoma (SL80) and the other grown in N. Carolina (SL81). SL80 and SL81 were stored at 2 to 5 C for 193 and 242 days, respectively. Samples were taken at about 28-day intervals for determination of germination, ethylene production, and LG activity. Seeds of two and three maturities were tested for SL80 and SL81, respectively. As afterripening of stored seeds proceeded, ethylene production gradually increased, with the maximum at 48 hours of germination. Germination showed a concomitant gradual increase. Lipoxygenase activity of both seedlots was less for mature than for immature seeds and showed a sharp increase during storage at 2 to 5 C, particularly for immature seeds. After heat-treatment to break dormancy of sublots from SL81, there was a progessive increase in ethylene production and germination, but most notably for mature seeds. In contrast to ethylene production and germination, after heat-treatment LG activity declined. Linear correlation coefficient (r) values between ethylene production and germination were highly significant for mature seeds from SL80 at 48 and 72 hours of germination, but only at 72 hours for immature seeds. For SL81 as for SL80, significant positive correlations were found between ethylene production and germination. However, correlations between LG activity and the other variables were not significant except for mature seeds from SL81. Significant positive correlations for both ethylene production and germination with LG activity also existed for these seeds. But after heat-treatment these correlations no longer occurred. The data indicate that the metabolic processes related to ethylene production and germination are occurring simultaneously with those of LG activity. The possibility that metabolites from LG activity serve as substrates for ethylene production can not be precluded.

Croissance et fructification de Ceratocystis spp. avec attention particulière à C. piceae et C. fimbriata, en fonction du substrat

1982 ◽  
Vol 60 (4) ◽  
pp. 358-363
Author(s):  
A. Thuillier ◽  
P. Neumann
Keyword(s):  
Nutrient Solution ◽  
Culture Media ◽  
Basal Medium ◽  
Malt Agar ◽  
Fruiting Bodies ◽  
Ceratocystis Fimbriata ◽  
Good Production ◽  
Wood Extracts ◽  
Standard Culture ◽  
Better Than

Ceratocystis coerulescens, C. fimbriata, C. ips, and C. minor were tested for production of sexual fruiting bodies, and C. penicillata and C. piceae for asexual fruiting bodies. Ceratocystis fimbriata produced perithecia easily on standard culture media, but there were marked differences between the two strains tested (503, 560). Strain 503 had a good production of fruiting bodies on malt agar (M) and a basal nutrient solution (N). Strain 560 fared better than 503 on Leonian agar (L), but did not fructify on M and N. Supplementing media with various wood extracts produced better results. M + maple sapwood extracts and L + poplar sapwood extracts gave the best results with strain 503, and L + pine sapwood extracts was the best with strain 560.Production of coremia was also influenced by the basal medium and the kind of extracts added as supplements. Fir and maple extracts stimulated the production of fruiting bodies, whereas pine and poplar extracts had no or very little stimulating effects. In every other species tested, the production of fruiting bodies was none or very irregular. [Journal translation]

scholarly journals In vitro answer of Bulgarian pepper (Capsicum annuum L.)

Genetika ◽  
2006 ◽  
Vol 38 (2) ◽  
pp. 129-136
Author(s):  
Velichka Rodeva ◽  
Stanislava Grozeva ◽  
Velichka Todorova
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Casein Hydrolysate ◽  
Basal Medium ◽  
Regeneration Ability ◽  
Stem Explants ◽  
Media Composition ◽  
Breeding Lines ◽  
High Level

Callusogenesis and regeneration ability of cotyledon and hypocotyl explants from three Bulgarian pepper varieties in MS basal medium supplemented with l-3mg/l BAP. l.0mg/1 IAA and 0.5mg/l GA3 was studied. In the different variants of culture medium was registered high level of callusogenesis and organogenesis in both type of explants from the all varieties. The highest percentage of plant-regenerants is established in cotyledon explants (from 3.3 to 18.3) in variant 3 of the culture medium containing 3mg/l BA. In the process of micropropagation by stem explants of the same studied pepper varieties the addition of the vitamins C. B12. Casein hydrolysate and Ferulic acid had a stimulation effect on the plant growth in height and rooting. In result of anther cultivation from three pepper varieties and four breeding lines the highest percentage of embryo structure formation was registered in varieties Albena and Strjama (12.0 and 13.8 respectively). The Bulgarian peppers are recalcitrant and their in vitro answer is different depending from the explants type, genotype and the culture media composition.

scholarly journals In vitro spore germination and early gametophyte development of Cibotium barometz (L.) J. Sm. in different media

2020 ◽  
Vol 21 (11) ◽  
Author(s):  
Yupi ISNAINI ◽  
Titien Ngatinem Praptosuwiryo
Keyword(s):  
Spore Germination ◽  
Culture Media ◽  
Basal Medium ◽  
Modern Medicine ◽  
Ex Situ ◽  
Naphthalene Acetic Acid ◽  
Gametophyte Development ◽  
In The Wild ◽  
Rare And Endangered

Abstract. Isnaini Y, Praptosuwiryo TNg. 2020. In vitro spore germination and early gametophyte development of Cibotium barometz (L.) J. Sm. in different media. Biodiversitas 21: 5373-5381. Cibotium barometz (L.) J. Sm. is known as the golden chicken fern and included in Appendix II of CITES. It is an important export commodity for traditional and modern medicine. Globally, populations of this species are under significant pressure due to overexploitation in the wild. In vitro culture is one of the technologies used for ex-situ propagation and conservation of rare and endangered ferns and lycophytes. This study’s objectives were: (i) to observe in vitro spore germination and early gametophyte development of C. barometz, and (ii) to determine the best culture medium for rapid spore germination and early development of the gametophytes. The sterilized spores were sown in half-strength Murashige & Skoog (½MS) basal medium supplemented with combinations of 6-Benzylaminopurine (BAP) and α-Naphthalene acetic acid (NAA). A factorial combination of four BAP concentrations (0, 2, 4, and 6 mg L-1) with four concentrations of NAA (0; 0.01; 0.03 and 0.05 mg L-1) created 16 treatments replicated in a Completely Randomized Design. Spore germination of C. barometz was observed to be Vittaria-type, and its prothallial development was Drynaria-type. Spore germination started 7-14 days after sowing. Young heart-shape gametophytes consisting of 110-240 cells were formed in 45-61 days after sowing. The two best spore culture media for rapid spore germination and development of C. barometz gametophytes were ½ MS with or without 2 mg L-1 BAP.

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