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Improvement of Straw Loading Method on Survival of Mouse IVF/IVC Blastocysts Cryopreserved by Vitrification (체외수정과 체외배양에서 생산된 생쥐 배반포기배의 초자화 동결에 있어서 Straw Loading 방법의 개선)

  • 김선의;엄상준;김은영;윤산현;박세필;임진호
    • Korean Journal of Animal Reproduction
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    • v.20 no.1
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    • pp.35-42
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    • 1996
  • This study was carried out to investigate the effect of straw loading method and thawing protocol on the in vitro development of in vitro produced mouse blastocysts cryopreserved by vitrification. Three loading types of straw I, Il and III on loading and sealing method were made for vitrification. The ability of the solution on straw loading methods to remain vitreous during warming was tested by exposed in air for 1 to 10 s sec. and then plunged the vitrified straws into water bath at 25°C. Embryos to be vitrified were equilibrated to the 20% EG for 5min. and exposed in EFS 40 for 1min. The plug ends of Straw I and Straw II were sealed with straw powder and Straw III was treated straw powder, followed by heat sealing and then plunged into LN$_2$. The results obtained in these experiments were summarized as follows; 1) Straw I embryo column mostly changed from transparent to opaque upon thawing without exposure in air for 3-6 sec. Straw II embryo column was I improved partially but was not remained completely vitreous during warming. However, when Straw lll loading method was used, the embryo column was remained vitreous completely. 2) High survival rates and development rates of each groups (middle blastocysts and hatching blastocysts) of vitrified embryos were obtained by using Straw III loading method than Straw I method (P<0.05). And the range of s standard error was low in Straw lll method. 3) When the embryos vitrified-frozen were placed in air for 3, 5 and l0sec. and then warmed rapidly in water bath at 25$^{\circ}C$, the survival rates after 24h of culture were 72.7-87.1% and the development rates to hatching stage after 48h of culture were 34.0-48.4%. There were no significantly differences according to exposure time in air during warming. In conclusion, the present results showed that highly survival and low standard error of vitrified-frozen mouse bIastocysts were obtained by using straw lll loading, double sealing and appropriate 2 step warming method.

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INFRARED THERMOGRAPHIC ANALYSIS OF TEMPERATURE RISE ON THE SURFACE OF BUCHANAN PLUGGER (적외선열화상장치를 이용한 Buchanan plugger 표면의 온도상승 분석)

  • Choi, Sung-A;Kim, Sun-Ho;Hwang, Yun-Chan;Youn, Chang;Oh, Byung-Ju;Choi, Bo-Young;Juhng, Woo-Nam;Jeong, Sun-Wa;Hwang, In-Nam;Oh, Won-Mann
    • Restorative Dentistry and Endodontics
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    • v.27 no.4
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    • pp.370-381
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    • 2002
  • This study was performed to evaluate the temperature rise on various position of the Buchanan plugger, the peak temperature of plugger's type and the temperature change by its touching time of heat control spling. The heat carrier system 'System B' (Model 1005, Analytic Technologies, USA) and the Buchanan's plug-gers of F, FM, M and ML sizes are used for this study. The temperature was set to 20$0^{\circ}C$ which Dr. Buchanan's "continuous wave of condensation" technique recommended on digital display and the power level on it was set to 10. In order to apply heat on the Buchanan's pluggers, the heat control spring was touched for 1, 2, 3, 4 and 5 seconds respectively. The temperature rise on the surface of the pluggers were measured at 0.5 mm intervals from tip to 20 mm length of shank using the infrared thermography (Radiation Thermometer-IR Temper, NEC San-ei Instruments, Ltd, Japan) and TH31-702 Data capture software program (NEC San-ei Instruments, Ltd, Japan). Data were analyzed using a one way ANOVA followed by Duncan's multiple range test and linear regression test. The results as follows. 1. The position at which temperature peaked was approximately at 0.5 mm to 1.5 mm far from the tip of Buchanan's pluggers (p<0.001). The temperature was constantly decreased toward the shank from the tip of it (p<0.001). 2. When the pluggerss were heated over 5 seconds, the peak temperature by time of measurement revealed from 253.3$\pm$10.5$^{\circ}C$ to 192.1$\pm$3.3$^{\circ}C$ in a touch for 1 sec, from 218.6$\pm$5.$0^{\circ}C$ to 179.5$\pm$4.2$^{\circ}C$ in a touch for 2 sec, from 197.5$\pm$3.$0^{\circ}C$ to 167.5$\pm$3.7$^{\circ}C$ in a touch for 3 sec, from 183.7$\pm$2.5$^{\circ}C$ to 159.8$\pm$3.6$^{\circ}C$ in a touch for 4 sec and from 164.9$\pm$2.$0^{\circ}C$ to 158.4$\pm$1.8$^{\circ}C$ in a touch for 5 sec. A touch for 1 sec showed the highest peak temperature, followed by, in descending order, 2 sec, 3 sec, 4 sec. A touch for 5 sec showed the lowest peak temperature (p<0.001). 3. A each type of pluggers showed different peak temperatures. The peak temperature was the highest in F type and followed by, in descending order, M type, ML type. FM type revealed the lowest peak temperature (p<0.001). The results of this study indicated that pluggers are designed to concentrate heat at around its tip, its actual temperature does not correlate well with the temperature which Buchanan's "continuous wave of condensation" technique recommend, and finally a quick touch of heat control spring for 1sec reveals the highest temperature rise.

Fine Structure of Mature Sperms of Cephalopods (Octopus minor, Octopus ocellatus and Todarodes pacificus) Inhabiting the Korean Waters II (한국 연근해산 두족류 (Octopus minor, Octopus ocellatus and Todarodes pacificus) 성숙정자의 미세구조 II)

  • Kim, Sang-Won;Chang, Nam-Sub
    • Applied Microscopy
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    • v.31 no.4
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    • pp.333-345
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    • 2001
  • The mature sperms of three species of cephalopods (Octopus minar, Octopus ocellatus, Todarodes pacificus) were observed by electron microscopy. The results obtained are as follows: The sperm lengths of Octopus minor and Octopus ocellatus of octopods are long and they are about $390{\mu}m$ and $125\sim130{\mu}m$, respectively, but the sperm length of Todarodes pacificus is short and about $35{\mu}m$. The sperm of Octopus minor has a helical acrosome and a head bent a little like a banana while Octopus ocellatus of octopod has a twisted acrosome and a long rod-shaped head. A number of horizontal stripes are observed as a periodic structure in their subacrosome cavities and dense plugs are formed in the cavities of their heads. On the other hand, the acrosome of Todarodes pacificus is circular cap-shaped, and its head is long and oval. It is notable that two small cavities were observed in its basal acrosome. Juxtanuclear acrosomal materials of high electron density filled the subacrosomal cavity. In the middle piece of mature sperms of Octopus minor and Octopus ocellatus, the mitochondria form the mitochondrial sleeve, but the numbers of mitochondria differ between the species so that they are $11\sim12$ and $8\sim9$, respectively. Meanwhile, in the middle piece of mature sperms of Todarodes pacificus, the mitochondria are separated from the axoneme, forming a mitochondrial spur in which $10\sim13$ mitochondria and some electron dense materials concentrate. The axoneme of Octopus minor, Octopus ocellatus and Todarodes pacificus are of 9+2 type in common, surrounded by 9 coarse fibres. A number of glycogen were observed only in the axoneme of Todarodes pacificus. The coarse fibres were found as far as the main piece of sperm tail in Octopks minor and Todarodes pacificus, while to the end piece of sperm tail in Octopus ocellatus.

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Design and Optimization of Pilot-Scale Bunsen Process in Sulfur-Iodine (SI) Cycle for Hydrogen Production (수소 생산을 위한 Sulfur-Iodine Cycle 분젠반응의 Pilot-Scale 공정 모델 개발 및 공정 최적화)

  • Park, Junkyu;Nam, KiJeon;Heo, SungKu;Lee, Jonggyu;Lee, In-Beum;Yoo, ChangKyoo
    • Korean Chemical Engineering Research
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    • v.58 no.2
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    • pp.235-247
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    • 2020
  • Simulation study and validation on 50 L/hr pilot-scale Bunsen process was carried out in order to investigate thermodynamics parameters, suitable reactor type, separator configuration, and the optimal conditions of reactors and separation. Sulfur-Iodine is thermochemical process using iodine and sulfur compounds for producing hydrogen from decomposition of water as net reaction. Understanding in phase separation and reaction of Bunsen Process is crucial since Bunsen Process acts as an intermediate process among three reactions. Electrolyte Non-Random Two-Liquid model is implemented in simulation as thermodynamic model. The simulation results are validated with the thermodynamic parameters and the 50 L/hr pilot-scale experimental data. The SO2 conversions of PFR and CSTR were compared as varying the temperature and reactor volume in order to investigate suitable type of reactor. Impurities in H2SO4 phase and HIX phase were investigated for 3-phase separator (vapor-liquid-liquid) and two 2-phase separators (vapor-liquid & liquid-liquid) in order to select separation configuration with better performance. The process optimization on reactor and phase separator is carried out to find the operating conditions and feed conditions that can reach the maximum SO2 conversion and the minimum H2SO4 impurities in HIX phase. For reactor optimization, the maximum 98% SO2 conversion was obtained with fixed iodine and water inlet flow rate when the diameter and length of PFR reactor are 0.20 m and 7.6m. Inlet water and iodine flow rate is reduced by 17% and 22% to reach the maximum 10% SO2 conversion with fixed temperature and PFR size (diameter: 3/8", length:3 m). When temperature (121℃) and PFR size (diameter: 0.2, length:7.6 m) are applied to the feed composition optimization, inlet water and iodine flow rate is reduced by 17% and 22% to reach the maximum 10% SO2 conversion.

Effects of Bisphenol A on the Placental Function and Reproduction in Rats (Bisphenol A가 흰쥐의 태반 기능과 출산에 미치는 영향)

  • Lee, Chae-Kwan;Kim, Seog-Hyun;Moon, Deog-Hwan;Kim, Jeong-Ho;Son, Byung-Chul;Kim, Dae-Hwan;Lee, Chang-Hee;Kim, Hwi-Dong;Kim, Jung-Won;Kim, Jong-Eun;Lee, Chae-Un
    • Journal of Preventive Medicine and Public Health
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    • v.38 no.3
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    • pp.330-336
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    • 2005
  • Objectives : The aim of this study was to investigate the effects of bisphenol A (BPA), an estrogen-like environmental endocrine disrupter, on the placental function and reproduction in rats. The mRNA levels of the placental prolactin-growth hormone(PRL-GH) gene family, placental trophoblast cell frequency and reproductive data were analyzed. Methods : The pregnancies of F344 Fisher rats ($160g{\pm}20g$) were detected by the presence of the copulatory plug or sperm in the vaginal smear, which marked Day 0 of pregnancy. Pregnant rats were divided into three groups. The control group was intraperitoneally injected with a sesame oil vehicle. The two remaining groups were injected with 50 or 500 mg/kg B.W/day of BPA, resuspended in sesame oil, on either days 7 to 11 or 16 to 20 of pregnancy, with the rats sacrificed on either day 11 or 20, respectively. The mRNA levels of PRL-GH and Pit-1a and b isotype genes were analyzed by Northern blot hybridization and reverse transcription-polymerase chain reaction. The hormone concentrations were analyzed by radioimmunoassay, and the frequency of the placental trophoblast cells observed by a histochemical study. Reproductive data, such as the placental weight and litter size, were surveyed on day 20. The fetal weight was surveyed for 4 weeks after birth. A statistical analysis was carried out using the SAS program (version 8.1). Results : The mRNA levels of the PRL-GH gene family, such as placental lactogen I, Iv and II, prolactin like protein A, C and Cv, and decidual prolactin-related protein were significantly reduced due to BPA exposure. The mRNA levels of the Pit-1a and b isotype genes, which induce the expression of the PRL-GH gene family in the rat placenta, were also reduced due to BPA exposure. The PL-Iv and PL-II concentrations were reduced in the BPA exposed group. During the middle to last stage of pregnancy (Days 11-20), a high dose of BPA exposure reduced the frequency of spongiotrophoblast cells, which are responsible for the secretion of the PRL-GH hormones. Reproductive data, such as the placental and fetal weights and the litter size, were reduced, but that of the pregnancy period was extended in the BPA exposed compared to the control group. Conclusions : BPA disrupts the placental functions in rats, which leads to reproductive disorders.

Effects of Chromium (VI) Exposure on the Placental Function and Reproduction in Rats (6가 크롬 폭로가 랫트의 태반 기능과 출산에 미치는 영향)

  • Lee, Heun;Moon, Deog-Hwan;Lee, Chae-Un;Kang, Sung-Goo;Son, Byung-Chul;Kim, Dae-Hwan;Lee, Chang-Hee;Kim, Jung-Won;Lee, Chae-Kwan;Chun, Jin-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.37 no.2
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    • pp.157-165
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    • 2004
  • Objectives : This study aimed to investigate the toxic effects of chromium (VI) on the placental function and reproduction in rats. For the study, the placental prolactin-growth hormone (PRL-GH) gene expression, placental trophoblast cell differentiation and reproductive data were analyzed. Methods : The pregnancies of F344 Fisher rats were checked by the presence of a copulatory plug or sperm in the vaginal smear, which was defined as day 0 of the pregnancy. Pregnant rats were divided into the three groups. The control group was given tap water (chromium level < 0.001 ppm) and the remaining groups were given 250 or 750 ppm of chromium (VI) [as potassium dichromate], from day 7 to 19 of the pregnancy. Rats were sacrificed at days 11 and 20 of pregnancy. The mRNA levels of PRL-GH and Pit-1a and b isotype genes were analyzed by Northern blot hybridization and reverse transcription-polymerase chain reaction (RT-PCR). The hormonal concentration was analyzed by radioimmunoassay, and the differentiation of placental trophoblast cells were observed by histochemical studies. Reproductive data, such as placental and fetal weights, pregnancy period, and litter size, were surveyed at day 20 of pregnancy and after birth. A statistical analysis was carried out using the SAS program (version 8.1). Results : The mRNA levels of the prolactin-growth hormone (PRL-GH) family of genes were dose dependently reduced by chromium exposure. The mRNA levels of Pit-1a and b isotype genes that induce the expression of the PRL-GH family of genes were also reduced by chromium exposure. The PRL-GH hormonal concentration in the rat placenta, fetus and maternal blood were decreased by chromium exposure. In the middle stage of pregnancy (day 11), a high dose of chromium suppressed the differentiation of spongiotrophoblast cells that secret the PRLGH hormones. In the last stage of pregnancy (day 20), a high dose of chromium induced apoptosis of placental cells. Reproductive data, such as placental and fetal weights, litter size, were reduced, but the pregnancy period was extended in the group exposed to chromium compared with the controls. Conclusion : Chromium (VI) disrupts the ordered functions of the placenta, which leads to reproductive disorders in rats.

Effect of Application Rate of a Controlled Release Fertilizer on the Changes in Medium EC and Growth of Subirrigated Vinca and Salvia (저면관수 재배에서 완효성 비료의 양이 배지의 EC 및 일일초와 살비아의 생장에 미치는 영향)

  • Kang, Jong Goo;Lee, In Ho
    • FLOWER RESEARCH JOURNAL
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    • v.16 no.2
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    • pp.112-117
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    • 2008
  • Plug seedlings of vinca (Catharanthus roseus L. 'Pacifica Punch') and salvia (Salvia splendens F. Sellow ex Roem & Schult 'Maestro') were transplanted into square plastic pots (145 mL volume) filled with a soilless growing medium. To determine the effect of application rate on the growing medium EC and growth of plants, 0, 0.5, 1.0, 1.5, 2.0 and 4.0 g per pot of a controlled release fertilizer (14-14-14 Osmocote, 14N-6.2P-11.6K) were mixed with the growing medium. Plants were subirrigated daily with tap water. In both vinca and salvia, growing medium EC increased as application rate was elevated. Growing medium EC was relatively constant over a whole crop period when the application rate was less than 1.5 g per pot, while it decreased throughout the experiment at higher application rates such as 2.0 to 4.0g per pot in both species. The greatest leaf area, plant height, and shoot dry weight of vinca were obtained when plants were fertilized with 2.0 to 4.0 g per pot of the fertilizer, resulting in a growing medium EC of $1.0{\sim}1.7dS{\cdot}m^{-1}$ throughout the experiment. Leaf area, shoot dry weight, and chlorophyll content of salvia increased with elevated application rates. Leaf area, shoot dry weight, and chlorophyll content of salvia were the greatest when plants were fertilized with 4.0 g per pot, resulting in growing medium EC of $1.0{\sim}4.0dS{\cdot}m^{-1}$ throughout the experiment. Plant height of salvia was the greatest when plants were fertilized with 2.0 to 4.0g per pot. Concentrations of nitrogen (N), phosphorus (P), potassium (K), magnesium (Mg), and sulfur (S) in the shoots of vinca increased, while concentration of calcium (Ca) decreased with elevated application rates. Concentrations of boron (B) and manganese (Mn) in the shoots of vinca increased as the application rate decreased.

Application of Chlorophyll Fluorescence Parameters for the Detection of Water Stress Ranges in Grafted Watermelon Seedlings (수박접목묘의 건조스트레스 범위 탐지를 위한 엽록소형광 지수의 적용)

  • Shin, Yu Kyeong;Kim, Yong Hyeon;Lee, Jun Gu
    • Journal of Bio-Environment Control
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    • v.28 no.4
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    • pp.461-470
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    • 2019
  • This study was carried out to quantify the drought stress in grafted watermelon seedlings non-destructively by using chlorophyll fluorescence (CF) imaging technique rather than the visual judgment. Six-day old watermelon seedlings were grown under uniform irrigation for 3 days, and then given drought stress. Afterward, the sensor for the measurement of water content in plug tray cell unit was used to classify the drought-stress level into nine groups from D1 (53.0%, sufficient moisture state) to D9 (15.7%, extremely dry stress), and the 16 CF parameters were measured. In addition, re-irrigation was performed on the drought stressed seedlings(D5 - D9) to determine the growth and photosynthesis recovery level, which was not confirmed by visual judgment. The kinetic curve patterns of CF in three different drought stressed seedling groups were found to be different for the early detection of drought stress. All the 16 CF parameters decreased continuously with exposure to drought stress and drastically decreased from D5 (32.1%) where the visual judgment was possible. The fluorescence decline ratio (Rfd_Lss) started to decrease from the initial drought stress level (D5 - D6), and the Maximum PSII quantum yield (Fv/Fm) was significantly decreased in the later extreme drought stress range (D7 - D9) by re-irrigation recovery test. Thus, Rfd_Lss and Fv/Fm parameters were finally selected as potent indicators of growth and photosynthesis recovery in the initial and later stages of drought stress. Also, to the differences in the numerical values of the individual chlorophyll fluorescence parameters, the drought stress level was intuitively confirmed through the image. These results indicate that Rfd and Fv/Fm can be considered as potential CF parameters for the detection of low and extremely high drought stress, respectively. Furthermore, Fv/Fm can be considered as the best CF parameters for recovery at re-irrigation.

Predictive Factors for Prognosis of Neonatal Intrahepatic Cholestasis : Non-Familial, Non-Metabolic, Non-Syndromic Cholestasis (신생아 간내 담즙 정체증의 예후 인자: 비가족성, 비대사성, 비증후성 담즙 정체증)

  • Kim, Hyung Suck;Lee, Chang Hoon;Kim, In Ju;Park, Jae Hong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.7 no.2
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    • pp.208-214
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    • 2004
  • Purpose: The prognosis of neonates with cholestasis is not clear. Some factors, such as high peak bilirubin levels and liver histologic findings have been claimed to affect the prognosis adversely. Our study aims to define which factors influence the prognosis of neonatal intrahepatic cholestasis. Methods: Retrospective reviews of the medical records were performed in 32 cases with neonatal intrahepatic cholestasis, who were admitted to Department of Pediatrics, Pusan National University Hospital from July 1995 to July 2002. Neonates were classified into 2 groups according to the duration of elevated serum alanine aminotransferase (ALT) levels more or less than 6 months. The data, such as biochemical, histopathologic and radiologic findings, were compared in both groups. Biochemical data included mean peak level of serum ALT, total bilirubin, direct bilirubin, and alkaline phosphatase. Histologic parameters related to lobular architecture, fibrosis, inflammatory infiltration and degenerative features of hepatocytes were arbitrary estimated on a scale of 1 to 3. Results: There were 19 males and 13 females, whose mean age was 48 days (14~77 days). The peak serum ALT levels were higher in the poor outcome group. Ductular proliferation and portoportal bridging were more severe in the poor outcome group. But the degree of multinucleated hepatocytes, hepatocellular swelling and canalicular plug did not appear to be significantly related to the long-term outcome. The DISIDA scintigraphy by visualization time of gall bladder and intestine was not useful in predicting outcome of neonatal intrahepatic cholestasis. Conclusion: Neonates who have intrahepatic cholestasis with high serum ALT levels, severe ductular proliferation and portoportal bridging in the liver biopsy specimen should be carefully followed up because they may have a poor prognosis.

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Enhanced Graft-take Ratio and Quality of Grafted Tomato Seedlings by Controlling Temperature and Humidity Conditions (토마토 공정묘의 접목활착율과 묘소질 향상을 위한 접목 활착실내의 적정 온.습도 조건 구명)

  • Vu, Ngoc-Thang;Zhang, Cheng-Hao;Xu, Zhi-Hao;Kim, Young-Shik;Kang, Ho-Min;Kim, Il-Soep
    • Journal of Bio-Environment Control
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    • v.22 no.2
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    • pp.146-153
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    • 2013
  • This study was conducted to enhance graft-take ratio and quality of grafted tomato seedlings by controlling temperature and humidity during the healing and acclimatization processes. Three temperature levels ($20^{\circ}C$, $23^{\circ}C$, and $26^{\circ}C$) were carried out to determine optimum temperature on four rootstocks. In addition, twelve combinations of three relative humidity levels (70%, 80%, and 90%) and four temperature levels ($17^{\circ}C$, $20^{\circ}C$, $23^{\circ}C$, and $26^{\circ}C$) were set up to evaluate the effect of relative humidity and temperature on the graft-take ratio of grafted seedlings. In the other hand, five relative humidity periods (H0, H1, H2, H3, and H4: 90% relative humidity for first 0, 1, 2, 3 and 10 days and afterwards relative humidity was reduced to 70%, respectively) were examined effect of relative humidity periods on the graft-take and quality of grafted seedlings. The higher graft-take ratios (84.0~87.4%) were showed at $23^{\circ}C$ compared to $20^{\circ}C$ and $26^{\circ}C$ in all rootstocks. Graft-take ratios decreased and number of diseased plants increased at high temperature. The graft-take ratios increased with increasing relative humidity in all temperature levels on the $3^{rd}$ and $7^{th}$ day after grafting. However, increasing relative humidity significantly increased percent of diseased plants. The graft-take ratio reduced at ($26^{\circ}C$) and ($17^{\circ}C$) temperature under all relative humidity conditions. The graft-take ratio increased with increasing period of 90% relative humidity. Maximum graft-take ratios were observed in H2 and H3 treatments. Graft-take ratio decreased with increasing 90% relative humidity for 10 days (H4). Diseased plants had not been found in H0, H1, H2, and H3 treatments. Seedling quality was improved through increasing fresh and dry weight of root, compactness, and root morphology of tomato seedlings in H2 and H3 treatments. Therefore, high relative humidity (90%) for first 2 or 3 days and afterwards reduced low relative humidity (70%) at $23^{\circ}C$ condition during healing and acclimatization promoted the graft-take and quality of grafted tomato seedlings.