• International Journal of Industrial Entomology and Biomaterials
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• v.11 no.2
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• pp.93-98
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• 2005

Studies on extended egg preservation schedule from 120 days to 180 days was taken up with 20 germplasm accessions of mutant silkworm genetic stocks of Bombyx mori L. Statistical analyses of the data collected over three trials revealed no significant changes both in the qualitative and quantitative traits of the genetic stocks between treatment (6 months egg preservation) and control (4 months egg preservation), except for fifth instar larval duration in TMS-61, TMS-62, TMS64, TMS-31 and TMS-34 shell weight in TMS-62, TMS-64 and TMS-66. Thus, the results indicate that extended schedule of 6 months egg preservation can safely be adopted, which will reduce the cost of conservation and minimize the genetic erosion owing to reduced crop cycle.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.1
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• pp.79-87
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• 2004

The silkworm rearing and growth parameters of 63 multivoltine silkworm accessions under extended period of egg preservation at 5$^{\circ}C$ from 30 days to 45 days were studied. The results indicate that, nine accessions did not respond to extended period of egg preservation at low temperature and the remaining 54 accessions responded to the treatment and three rearings were conducted for comparision with the control; to estimate the effect of prolonged egg preservation at low temperature. The non-parametric tests statistics (Wilcoxon tests) was adopted for comparing the mean performance of treated batches (45 days) over the control (30 days). Highly significant variability was found among the accessions for all the parameters under study. The genetically controlled morphological characters were not altered in the treated batches, which were found to be on par with that of control. However, the total larval duration varied significantly over the control in 51 accessions. Similarly, the fifth age larval duration of 27 accessions showed decreasing trend compared to control. Altogether 41 accessions were found to be tolerant to long-term cold preservation upto 45 days, without showing any significant variation for morphological as well as essential quantitative traits. These accessions may be recommended for long-term egg preservation schedule up to 45 days, which will reduce the cost of conservation of these silkworm germplasm.

• International Journal of Industrial Entomology and Biomaterials
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• v.15 no.1
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• pp.1-7
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• 2007

At present, multivoltine silkworm races reared five times per year involving huge manpower and rearing expenditure. Egg diapausing behavior is facultative in multivoltine and egg diapause was induced in selected multivoltine races by regulating temperature at $18^{\circ}C$, relative humidity 80% and photoperiod (6L:18D) in the late stage silkworm rearing. The maximum percentage of egg diapause induction was recorded in Rong Diazo, Diazo and MW13 showed 94%, 93% and 92% respectively, whereas the races A14DY and OS-616 showed minimum diapause induction 15% and 18% respectively. The diapause induced multivoltine eggs were preserved up to six months by cold preservation schedule normally adopted for bivoltine. After three and six months egg preservation, the diapause induced layings were released and observed for hatching percentage, all races showed above 82 % of hatching except the race AP12, which showed only 78 % of hatching. This methods reduce the crop cycle, gives strong safety backup and preventing the genetic erosion. This study helps formulating a new conservation method for multivoltine silkworm germplasm.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.35-35
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• 2022

Pears (Pyrus spp.) have been grown worldwide as a kind of important economical fruits. Over 1,500 accessions collected from countries have been preserved in National Institute of Horticultural and Herbal Science, Rural Development Administration in Korea. However, redundancies and misidentification are happening in the germplasm preservation due to same cultivars which have different names in various localities (synonyms) and different cultivars which have same names (homonyms). That can lower germplasm management efficiency. The object of this study is to identify synonyms and homonyms in pear germplasms by analyzing genetic variation with four microsatellite markers: CH03d12, CH03g07, CH02b10, and EMPc117. PCR amplification with above 4 microsatellite markers was done for the 31 pear accessions, and the products were analyzed by agarose gel electrophoresis. As a result, 7 synonyms and 9 homonyms were identified among 31 pear accessions. We'll compare these genotypes with phenotypes of each pear accessions, and reduces the redundancy and misidentification in pear germplasm collection for the reliable management.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.7
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• pp.939-945
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• 2003

Seventeen Chinese indigenous pig breeds and three introduced pig breeds had been carried out by means of vertical polyacrylamide gel electrophoresis (PAGE). According to the results, eight serum protein loci were highly polymorphic except Pi-2 and Cp. The polymorphism information content (PIC) of Hpx was the highest (0.5268), while that of Cp was the lowest (0.0257). The population genetic variation index showed that about 84% genetic variation existed in the population, and the rest of 16% distributed between the populations. The genetic variation of Yimeng black pig and Duroc were the highest and the lowest, respectively. The genetic variation of Chinese indigenous pig breeds was much more than that of exotic groups. Genetic distance results showed that Chinese indigenous pig breeds were classified into four groups with the three introduced pig breeds clustered into another group. The results also supported the geographic distribution of Chinese indigenous pig breeds in certain extent.

• Korean Journal of Plant Resources
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• v.30 no.6
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• pp.612-617
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• 2017

The preservation of pear germplasm, like that of other clonal germplasms, is difficult because it requires conservation of whole plants or their tissues. Among the currently available methods for long-term conservation of clonal germplasm, cryopreservation of shoot tips is the most reliable and cost- and space-effective option. Alginate-coated axillary shoot tips from in vitro-grown pear were conserved successfully in liquid nitrogen (LN) following dehydration. Shoot recovery from cryopreserved shoot tips was improved greatly after 8 weeks of cold acclimation, but recovery decreased slightly after then. The highest regeneration rate was observed when in vitro shoot tips were preincubated in MS (Murashige and Skoog) medium with 0.3 M sucrose for 48 h, and when alginate-coated shoot tips were precultured in MS medium with increasing sucrose concentrations (0.5 M and 0.7 M) for 8 and 16 h, respectively. When the encapsulated beads were dehydrated for up to 7 h [25% water content (fresh weight basis)] under laminar flow, the highest regeneration rate was observed in "BaeYun No. 3" (55.7%) and "Whanggeum" (43.3%) after warming from LN. This technique is useful as a practical procedure to cryopreserve plant material that is sensitive to freezing of the surrounding cryoprotectant medium. Therefore, this technique appears to be promising for the cryopreservation of shoot tips from in vitro-grown plantlets of pear germplasm.

• Interdisciplinary Bio Central
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• v.2 no.4
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• pp.16.1-16.4
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• 2010

Cryopreservation of mouse embryos and spermatozoa has become the foremost technique for preserving large numbers of different strains of mice with induced mutations. In 1998, our mouse bank was established in the Center for Animal Resources and Development (CARD), Institute of Resource Development and Analysis, Kumamoto University, Japan, based on the Preservation, supply and development of genetically engineered animals report published by the Ministry of Education, Culture, Sports, Science and Technology. We cryopreserve mouse embryos and sperm, supply these resources, organize training courses to educate people and form part of a domestic and international network of both mutagenesis and resource centers. We currently have over 1,500 mouse strains, 842,000 frozen embryos and 26,000 straws containing frozen sperm. Moreover, we disclose information about 1,300 deposited strains. Furthermore, over 400 strains of frozen embryos or mice produced from frozen embryos and sperm are being supplied to the requesters both domestically and internationally. Additionally we hold training courses on the cryopreservation of mouse germplasm 2~3 times a year, both domestically and internationally. In the course, we teach basic reproductive engineering techniques to trainees on a man-to-man basis. We have already held 28 training courses on the cryopreservation of mouse germplasm at our center and at other institutes.

• Korean Journal of Plant Resources
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• v.36 no.6
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• pp.562-570
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• 2023

The droplet-vitrification technique for cryopreservation has proven successful across a diverse range of germplasm, ensuring safe and effective long term preservation. In this study, we investigate an effective cryopreservation protocol using the droplet-vitrification technique for shoot tips of Freesia hybrida cultivars 'Sunny Gold' and 'Sweet Lemon'. To determine optimal conditions for Freesia cryopreservation, we employed a carefully selected standard procedure along with additional treatments and alternative solutions. For 'Sunny Gold', the highest regrowth rate of 24% was achieved when shoot tips underwent dehydration with PVS3 solution for 120 minutes before direct immersion in liquid nitrogen (LN) for 1 hour, coupled with a standard protocol involving a two-step preculture with 0.3 M - 0.5 M sucrose, loading with C4 for 40 minutes, and unloading with 0.8 M sucrose for 40 minutes. In the case of 'Sweet Lemon,' regrowth of cryopreserved shoot tips was observed with dehydration treatments, including PVS2 (A3) for 60 minutes and PVS3 (B1) for 60 minutes, as well as longer exposure. The results reflect the distinct sensitivity of shoot tips to chemical toxicity and osmotic stress in these two genotypes. This study provides valuable evidence to consistently enhance the effectiveness of cryopreservation methods for the long-term conservation of Freesia germplasm.

• Journal of Sericultural and Entomological Science
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• v.42 no.1
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• pp.1-5
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• 2000

To investigate the possibility of crypreservation of mulberry seeds in liquid nitrogen(LN$_2$), characteristics of the seeds were examined after picking mulberry syncarps and drying-heat treatment. Storage in LN$_2$has the potential of providing indifinite preservation of valuable seed germplasm. Determining the tolerance of seeds among given cultivars to LN$_2$cooling and subsequent rewarming is the first step to establishing the feasibility of LN$_2$storage. Seeds of 4 mulberry varities were treated to LN$_2$(-196$\^{C}$) for 24 hours after drying heat treatment. Seed moisture content of Daeryukppong was the highest. As moisture content of mulberry seed was below 1%, storage in LN$_2$was safe. And drying heat treatment for 60 minutes was suitable to prevent decreased germination rate and germination vigor of seeds. The seeds of Cheongilppong were unsuibable to cryopreserve in LN$_2$for longterm storage.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.11
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• pp.1527-1533
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• 2001

Garole is a prolific, rare, less known and small size Indian sheep breed found in low and humid Sunderban region of West Bengal. Although information on stored Garole ram liquid semen upto 24 h is available, but there is a need to further investigate the short-term and long-term preservability of Garole ram semen for extensive utilization of this valuable germplasm by artificial insemination. The aim of the present study was to apply computer-assisted sperm analysis technique for assessing the motion characteristics of Garole ram semen stored (i) in liquid state at refrigeration temperature for short-term preservation upto 48 h and (ii) in frozen state at $-196^{\circ}C$ for long-term preservation after packaging in mini straws. Short-term preservation had a significant effect on motility (p<0.01) as the motility progressively decreased from 90.1% at 0 h to 85.5% and 73.2% after 24 and 48 h of storage, respectively. Although the decline in rapid moving sperms was also significant (p<0.01) on storage but the decrease was more pronounced at 48 h as compared to 24 h of storage period. Storage of chilled semen had also a significant effect on % linearity (p<0.05), % straightness (p<0.01), sperm velocities (p<0.01), amplitude of lateral head displacement (p<0.01) and beat frequency (pO.Ol) of spermatozoa. The replication had a significant effect for all the variables except average path and straight line velocity. However, the interactions of short-term storage and replication were non-significant for most of the variables except % of medium moving sperms, sperm velocities and beat frequency. On long-term preservation of Garole ram spermatozoa under controlled conditions the mean post-thaw recovery of 70.4 and 71.4% motile spermatozoa was achieved having 48.8 and 48.9% of rapidly motile spermatozoa, respectively in both the replicates. The effect of replication on cryopreservation was significant (p<0.05) on amplitude of lateral head displacement and beat frequency, but there was no significant effect on motility, rapidly motile spermatozoa, linearity, straightness and sperm velocities of frozen-thawed spermatozoa. It can be concluded from these results that an average 70% motility can be achieved on storage of Garole ram semen in chilled liquid state upto 48 h or in liquid nitrogen after freezing under controlled conditions in straws. However, further studies are required to evaluate the fertility of short-term and long-term preserved Garole ram semen for extensive use of this prolific sheep breed.