• Title/Summary/Keyword: 저항성 유전

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High-efficiency development of herbicide-resistant transgenic lilies via an Agrobacterium-mediated transformation system (고효율의 아그로박테리움 형질전환법을 이용한 제초제저항성 나리 식물체 개발)

  • Jong Bo Kim
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.56-62
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    • 2023
  • Transgenic lilies have been obtained using Agrobacterium tumefaciens (AGL1) with the plant scale explants, followed by DL-phosphinothricin (PPT) selection. In this study, scales of lily plants cv. "red flame" were transformed with the pCAMBIA3301 vector containing the gus gene as a reporter and the blpR gene as a selectable marker, as well as a gene of interest showing herbicide tolerance, both driven by the CaMV 35S promoter. Using a 20-minute infection time and a 5-day cultivation period, factors that optimized and demonstrated a high transformation efficiency were achieved. With these conditions, approximately 22-27% efficiency was observed for Agrobacterium-mediated transformation in lilies. After transformation with Agrobacterium, scales of lilies were transferred to MS medium without selective agents for 2 weeks. They were then placed on selection MS medium containing 5 mg/L PPT for a month of further selection and then cultured for another 4-8 weeks with a 4-week subculture regime on the same selection medium. PPT-resistant scales with shoots were successfully rooted and regenerated into plantlets after transferring into hormone-free MS medium. Also, most survived putatively transformed plantlets indicated the presence of the blpR gene by PCR analysis and showed a blue color indicating expression of the gus gene. In conclusion, when 100 scales of lily cv. "red flame" are transformed with Agrobacterium, approximately 22-27 transgenic plantlets can be produced following an optimized protocol. Therefore, this protocol can contribute to the lily breeding program in the future.

A New Medium Maturing and High Quality Rice Variety with Lodging and Disease Resistance, 'Jinbo' (중생 고품질 내도복 내병성 벼 품종 '진보')

  • Kim, Jeong-Il;Park, No-Bong;Lee, Ji-Yoon;Park, Dong-Soo;Yeo, Un-Sang;Chang, Jae-Ki;Kang, Jung-Hun;Oh, Byeong-Geun;Kwon, Oh-Deog;Kwak, Do-Yeon;Lee, Jong-Hee;Yi, Gi-Hwan;Kim, Chun-Song;Song, You-Cheon;Cho, Jun-Hyun;Nam, Min-Hee;Choung, Jin-Il;Shin, Mun-Sik;Jeon, Myeong-Gi;Yang, Sae-Jun;Kang, Hang-Weon;Ahn, Jin-Gon;Kim, Jae-Kyu
    • Korean Journal of Breeding Science
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    • v.43 no.3
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    • pp.165-171
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    • 2011
  • A new rice variety 'Jinbo' is a japonica rice (Oryza sativa L.) with good eating quality, lodging tolerance, and resistance to rice stripe virus (RSV) and bacterial blight disease (BB). It was developed by the rice breeding team of Yeongdeog Substation, National Institute of Crop Science (NICS), RDA in 2009. This variety was derived from a cross between 'Yeongdeog26' with good grain quality and wind tolerance and 'Koshihikari' with good eating quality in 1998 summer season. A promising line, YR21324-56-1-1, selected by pedigree breeding method, was designated as the name of 'Yeongdeog45' in 2005. After the local adaptability test was carried out at nine locations from 2006 to 2008, 'Yeongdeog45' was released as the name of 'Jinbo' in 2009. 'Jinbo' has short culm length as 74 cm and medium maturating growth duration. This variety is resistant to $K_1$, $K_2$, and $K_3$ races of bacterial blight and stripe virus and moderately resistant to leaf blast disease with durable resistance, and also it has tolerance to unfavorable environments such as cold and dried wind. 'Jinbo' has translucent and clear milled rice kernel without white core and white belly rice, and good eating quality as a result of panel test. The yield potential of 'Jinbo' in milled rice is about 5.65 MT/ha at ordinary fertilizer level in local adaptability test. This cultivar would be adaptable to middle plain, mid-west costal area, east-south coastal area, and south mid-mountainous area.

Evaluation of $SrRuO_3$ Buffer Layer for $Pb(Zr,Ti)O_3$ Ferroelectric Capacitor ($Pb(Zr,Ti)O_3$ 강유전체 커패시터에 적용하기 위한 $SrRuO_3$ 버퍼 층의 특성 평가)

  • Kweon, Soon-Yong;Choi, Ji-Hye;Son, Young-Jin;Hong, Suk-Kyoung;Ryu, Sung-Lim
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2007.06a
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    • pp.280-280
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    • 2007
  • $Pb(Zr,Ti)O_3$ (PZT) 강유전체 박막은 높은 잔류 분극 (remanent polarization) 특성 때문에 현재 강유전체 메모리 (FeRAM) 소자에 적용하기 위하여 가장 활발히 연구되고 있다. 그런데 PZT 물질은 피로 (fatigue) 및 임프린트 (imprint) 등의 장시간 신뢰성 (long-term reliability) 특성이 취약한 단점을 가지고 있다. 이러한 신뢰성 문제를 해결할 수 있는 효과적인 방법 중의 하나는 $IrO_2$, $SrRuO_3$(SRO) 등의 산화물 전극을 사용하는 것이다. 많은 산화물 전극 중에서 SRO는 PZT와 비슷한 pseudo-perovskite 결정구조를 갖고 격자 상수도 비슷하여, PZT 커패시터의 강유전 특성 및 신뢰성을 향상시키는데 매우 효과적인 것으로 알려져 있다. 따라서 본 연구는 PZT 커패시터에 적용하기 위하여 SRO 박막을 증착하고 이의 전기적 특성 및 미세구조를 분석하고자 하였다. 또 실제로 SRO 박막을 상부전극과 PZT 사이의 버퍼 층 (buffer layer)으로 적용한 경우의 커패시터 특성도 평가하였다. 먼저 다결정 SRO 박막을 $SiO_2$/Si 기판 위에 DC 마그네트론 스퍼터링 법 (DC magnetron sputtering method)으로 증착하였다. 그 다음 이러한 SRO 박막의 미세구조, 결정성 및 전기적 특성이 증착 조건들의 변화에 따라서 어떤 경향성을 보이는지를 평가하였다. 기판 온도는 $350\;{\sim}\;650^{\circ}C$ 범위에서 변화시켰고, 증착 파워는 500 ~ 800 W 범위에서 변화시켰다. 또 Ar+$O_2$ 혼합 가스에서 산소의 혼합 비율을 20 ~ 50% 범위에서 변화시켰다. 이러한 실험 결과 SRO 박막의 전기적 특성 및 미세 구조는 기판의 증착 온도에 따라서 가장 민감하게 변함을 관찰할 수 있었다. 다른 증착 조건과 무관하게 $450^{\circ}C$ 이상의 온도에서 증착된 SRO 박막은 모두 주상정 구조 (columnar structure)를 형성하며 (110) 방향성을 강하게 나타내었다. 가장 낮은 전기 저항은 $550^{\circ}C$ 증착 온도에서 얻을 수 있었는데, 그 값은 약 $440\;{\mu}{\Omega}{\cdot}cm$ 이었다. SRO 버퍼 충을 적용하여 제작한 PZT 커패시터의 잔류 분극 (Pr) 값은 약 $30\;{\mu}C/cm^2$ 정도로 매우 높은 값을 나타내었고, 피로 손실 (fatigue loss)도 $1{\times}10^{11}$ 스위칭 사이클 후에 약 11% 정도로 매우 양호한 값을 나타내었다.

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Study on the Genetic Characteristics of Waterlogging Tolerant Pepper (Capsicum annuum L.) for Breeding Tolerant Varieties against Flooding Stress (내습성 고추 품종 육성을 위한 선발계통의 유전적 특성 구명)

  • Yang, Eun Young;Chae, Soo-Young;Hong, Jong-Pil;Lee, Hye-Eun;Park, Eun Joon;Moon, Ji-hye;Park, Tae-Sung;Roh, Mi-Young;Kim, Ok Rye;Kim, Sang Gyu;Kim, Dae Young;Lee, Sun Yi;Cho, Myeong Cheoul
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1111-1120
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    • 2017
  • This study was conducted to select pepper lines that were tolerant to excessive water injury among the pepper germplasm and investigate the genetic characteristics of those lines to contribute to the breeding of pepper cultivars with stable productivity in abnormal weather. Each of the tolerant and susceptible lines went through immersion treatment, and differentially expressed genes between them were analyzed. The tolerant line showed increased expression of the CA02g26670 gene, which is involved in the CONSTANS protein pathway and regulation of flowering by day length, but it exhibited decreased expressions of CA01g21450, CA01g22480, CA01g34470, CA02g00370 and CA02g00380. The susceptible line showed increased gene expressions of CA02g09720, CA02g21290, CA03g16520, CA07g 02110, and CA12g17910, which are involved in the inhibition of proteolytic enzyme activity, DNA binding, inhibition of cell wall-degrading enzyme, and inhibition of nodulin, respectively. Meanwhile the expressions of CA02g02820, CA03g21390, CA06g17700 and CA07g18230 decreased in the susceptible line, in relation to calcium-ion binding, high temperature, synthesis of phosphocholine and cold stress, respectively. The expressions of genes related to apoptosis and peroxidase increased, while that of CA02g16990, which functions as a nucleoside transporter, decreased in both the tolerant and susceptible lines. Based on the different gene expressions between the tolerant and susceptible lines, further studies are needed on breeding abiotic stress-tolerant lines.

Selection of Virulent Isolates of Fusarium circinatum and Investigation of Pitch Canker Severity of Pinus rigida and P. rigida × P. taeda Seed Orchards in Jeju Island (제주도(濟州道) 리기다 및 리기테다 채종원(採種園)에서 푸사리움가지마름병 피해도(被害度) 조사(調査) 및 병원성(病原性) 균주선발(菌株選拔))

  • Woo, Kwan-Soo;Kim, Young-Joung;Kim, Tae-Su;Lee, Seong-Kyu
    • Journal of Korean Society of Forest Science
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    • v.94 no.6
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    • pp.402-409
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    • 2005
  • This study was undertaken to compare and estimate the severity of pitch canker of individual trees of Pinus rigida and Pinus rigida ${\times}$ P. taeda in two seed orchards in Jeju island, in which the orchards have been damaged by the pitch canker for seven years. Wind-pollinated two-year-old seedlings of P. rigida and P. rigida ${\times}$ P. taeda, in which the seedlings of P. rigida ${\times}$ P. taeda were from seeds of phenotypically selected, uninfected(but untested) trees, were inoculated with the pathogenic fungus, Fusarium circinatum, isolated from P. rigida and P. thunbergii. The virulence of the isolates was also identified. Statistically significant difference was found in 'stem cankers'(SC; ${\chi}^2=7.76$, P=0.05) among 4 plantations of P. rigida ${\times}$ P. taeda of two seed orchards. P. rigida was higher in 'top kill' (TK) and 'branch tip symptoms' (BT) than those of P. rigida ${\times}$ P. taeda. In artificial inoculation tests, mortality of the seedlings from the resistant candidates was 14% higher than that of the seedlings from the susceptible candidates. This result may becaused by unknown pollen trees and/or candidate tree selection based only on phenotype. Two of five fungal isolates, C-6-L(9) and C-6-L(19), showed significantly higher mortality (68% and 60%, respectively) than others, suggesting that these isolates can be used as virulent isolates for a mass artificial inoculation. Resistance candidate seedlings that were selected from this study can be utilized as useful materials for fundamental studies of genetics and biochemistry to breed resistance varieties to pitch canker.

Biochemical Characterizations of Phenylalanine Ammonia-Lyase and its Mutants to Develop an Enzymatic Therapy for Phenylketonuria (페닐케톤뇨증의 효소치료 개발을 위한 phenylalanine ammonia-lyase 및 유전자 변이형의 생화학적 특성)

  • Kim, Woo-Mi
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1226-1231
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    • 2009
  • Enzyme substitution with recombinant phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is currently being explored for treatment of phenylketonuria (PKU), an autosomal recessive genetic disorder with mutations of the gene encoding phenylalanine-4-hydroxylase (EC 1.14.16.1). However, oral administration of PAL is limited because of proteolytic digestion in the gastrointestinal tract. The aim of this study was to determine the biochemical properties of PAL and delinate the susceptibility of wild-type PAL to pancreatic proteolysis by exploring several mutants, and to develop therapeutic drugs with PAL for PKU. The specific activity of PAL was assayed and its optimal pH, temperature stability, and intestinal protease susceptibility were investigated. Its $V_{max}$ values for phenylalanine and tyrosine were 1.77 and $0.47{\mu}mol$/ min/mg protein, respectively, and its $K_m$ values were $4.77{\times}10^{-4}$ and $4.37{\times}10^{-4}\;M$, respectively. PAL showed an optimal pH at 8.5, corresponding to the average pH range of the small intestine. It showed no loss of activity at $-80^{\circ}C$ for 5 months and possessed 93.4% of its activity under $4^{\circ}C$ for 4 wks. PAL was susceptible to chymotrypsin digestion and, to a lesser extent, to trypsin, elastase, carboxypeptidase A, and B. The trypsin and chymotrypsin cleaving sites were mutated to investigate protection from pancreatic digestion and the specific activities of these mutants were evaluated. The six mutants displayed low specific activities compared to the wild-type, suggesting that the primary trypsin and chymotrypsin cleaving sites may be essential for catalytic reaction. The PAL mutants could therefore be applied as a pretreatment modality without susceptibility to proteolytic attack, however, additional modification for enhancing enzymatic activity is needed to reduce the Phe levels effectively.

A Study on Batch-Type Remote Plasma Dry Cleaning Process for Native Oxide Removal (배치식 플라즈마 세정 설비를 이용한 자연산화막 제거 공정)

  • Park, Jae-Young;Yi, Wook-Yeol;Hyung, Yong-Woo;Nam, Seok-Woo;Lee, Hyeon-Deok;Song, Chang-Lyong;Kang, Ho-Kyu;Roh, Yong-Han
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2004.11a
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    • pp.247-251
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    • 2004
  • 반도체 소자의 제조에 있어 실리콘 표면에 성장한 자연산화막을 제거하기 위해 일반적으로 습식 세정 기술이 이용되어 왔다. 하지만 소자의 최소 선폭(design rule)이 nano급으로 고집적화 됨에 따라 contact hole 바닥의 자연산화막을 깨끗이 제거하는데 있어서 그 한계를 나타나고 있다. 이에 대한 효과적인 대안 공정으로 가스 건식 세정 기술이 연구되고 있다. 본 논문에서는 한 번에 50매 이상의 웨이퍼를 처리함으로써 생산성 측면에서 월등한 배치식 설비에서 원거리 플라즈마(remote plasma) 장치에서 2.450Hz의 마이크로웨이브(${\mu}$-wave)에 의해 형성시킨 수소라디칼과 $NF_3$ 가스를 이용하여 실리콘에 결함을 주지 않고 자연산화막을 선택적으로 제거하는 공정에 대해 고찰하였다. AFM을 이용한 표면분석, TEM을 이용한 물성분석, 그리고 ToF-SIMS 및 XPS를 이용한 화학 분석을 습식 및 건식 세정을 비교 평가한 결과, 건식 세정 공정이 실리콘 표면에 결함을 주지 않고 자연산화막을 제거 할 수 있음을 확인하였다. 산화막$(SiO_2)$, 질화막$(Si_3N_4)$, 그리고 다결정 실리콘(Poly-Si) 등의 각 막질별 식각 특성을 고찰하였으며, $NH_3$의 캐리어 가스인 $N_2$의 주입량을 조절함으로써 수소라디칼 형성 효율의 개선이 가능하였으며, 이로부터 게이트와 소스/드레인 사이를 절연하기 위해 이용되는 질화막의 식각 선택비를 2배 정도 개선할 수 있었다. nano급 소자에 실장하여 평가한 결과에서 불산(HF)에 의한 습식 세정 방식에 비하여 약 $20{\sim}50%$ 정도의 contact 저항 감소 효과가 있음이 확인되었다.두 소자 모두 $40mA/cm^2$ 에서 이상적인 화이트 발란스와 같은(0.33,0.33)의 색좌표를 보였다.epsilon}_0=1345$의 빼어난 압전 및 유전특성과 $330^{\circ}C$의 높은 $T_c$를 보였고 그 조성의 vibration velocity는 약4.5 m/s로 나타났다.한 관심이 높아지고 있다. 그러나 고 자장 영상에서의 rf field 에 의한 SAR 증가는 중요한 제한 요소로 부각되고 있다. 나선주사영상은 SAR 문제가 근원적으로 발생하지 않고, EPI에 비하여 하드웨어 요구 조건이 낮아 고 자장에서의 고속영상방법으로 적합하다. 본 논문에서는 고차 shimming 을 통하여 불균일도를 개선하고, single shot 과 interleaving 을 적용한 multi-shot 나선주사영상 기법으로 $100{\times}100$에서 $256{\times}256$의 고해상도 영상을 얻어 고 자장에서 초고속영상기법으로 다양한 적용 가능성을 보였다. 연구에서 연구된 $[^{18}F]F_2$가스는 친핵성 치환반응으로 방사성동위원소를 도입하기 어려운 다양한 방사성의 약품개발에 유용하게 이용될 수 있을 것이다.었으나 움직임 보정 후 영상을 이용하여 비교한 경우, 결합능 변화가 선조체 영역에서 국한되어 나타나며 그 유의성이 움직임 보정 전에 비하여 낮음을 알 수 있었다. 결론: 뇌활성화 과제 수행시에 동반되는 피험자의 머리 움직임에 의하여 도파민 유리가 과대평가되었으며 이는 이 연구에서 제안한 영상정합을 이용한 움직임 보정기법에 의해서 개선되었다. 답이 없는 문제, 문제 만들기, 일반화가 가능한 문제 등으로 보고, 수학적 창의성 중 특히 확산적 사고에 초점을 맞추어 개방형 문제가 확

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BCR (Brown Color Repressor) gene isolation related to mycelial browning of Lentinus edodes (표고균사 갈변과 관련된 BCR (Brown Color Repressor) 유전자 분리)

  • Kim, Young-Ho;Park, Soo-Cheol;Jhune, Chang-Sung;You, Chang-Hyun;Sung, Jae-Mo;Kong, Won-Sik
    • Journal of Mushroom
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    • v.10 no.3
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    • pp.120-128
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    • 2012
  • Recently sawdust cultivation of Shiitake mushroom (Lentinula edodes) is increasing. It is important to make mycelia to be brown on the substrate surface. This browned surface in sawdust cultivation plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. In order to isolate genes which related to brown color formation, differential display method was used. Two cDNA fragments obtained by DD-PCR were 1.2 and 1.6kb and these were expressed in white colored mycelia from L. edodes, but not brown colored mycelia. Partial sequencing of these cDNA fragments showed that the 1.6kb cDNA had 100% identity with the microsatellites gene from Dugenia polichroa. However, the other 1.2kb cDNA fragment had poly T tail on 3' region of partial open reading frame on 5' region. The new primer designed based on the sequence of 1.2kb cDNA was constructed. RT-PCR analysis using the newly designed 0.12kb cDNA specific primer showed that the gene was only expressed in white color mycelia, but not in brown color mycelia. Sequence analysis of 5' region of this 1.2kb cDNA revealed that this gene contained partial open reading frame consisted of 110 amino acid. Homology search using DNASIS database showed that this gene had high sequence homology of 66.7% in DNA level and 69.2 % in amino acid level with dTDP-glucose 4,6-dehydratases gene from Arabidopsis thaliata. The dTDP-glucose 4,6-dehydratases gene was known to be function to have tolerance with oxidation stress. These results strongly suggest that this gene isolated from white mycelia of L. edodes might have a function of repressor against mycelia browning. Therefore I designated this gene as BCR (Brown Color Repressor) gene.

Investigation of Transformation Efficiency of Rice Using Agrobacterium tumefaciens and High Transformation of GPAT (glycerol-3-phosphate acyltransferase) Gene Relative to Chilling Tolerance (Agrobacterium tumefaciens를 이용한 벼의 형질전환 효율의 검토 및 내한성 관련 GPAT (glycerol-3-phosphate acyltransferase) 유전자의 형질전환)

  • Seo, Mi-Suk;Bae, Chang-Hyu;Choi, Dae-Ock;Rhim, Seong-Lyul;Seo, Suk-Chul;Song, Pill-Soon;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.85-92
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    • 2002
  • This study has been focused on improving transformation efficiency of rice using Agrobacterium tumefaciens. We have demonstrated the effect of this system when the GPAT gene related to the cold-resistance was transferred by Agrobacterium tumefaciens in rice. Transformation conditions were modified using intron $\beta$-glucuronidase (GUS) expression as a reporter gene in the rice. In this study, mature seed-derived calli of rice (Oruza sativa L. cv. Dongjin) were pre-cultured for 3 days and then infected with Agrobacterium. When this infected calli were cultured in the dark for 10 days on co-cu]lure medium containing 50 mg/L of CaCl$_2$, 30 mg/L of acetosyringone, 2 mg/L of 2,4-D, 120 mg/L of betaine, high GUS expression was observed. In the present transformation system, the efficiency of transformation of GPAT gene was about 54%. Stable integration of GPAT gene into chromosomal DNA was proven by southern blot analysis of genomic DNA isolated from T$_{0}$ progenies. The progenies (T1 generation) derived from primary transformant of 5 lines were segregated with a 3 (resistant) : 1 (sensitive ratio) in medium containing hygromycin. This high frequency transformation system can be used as a useful tool in transformation of another monocotyledon.n.

Floral Bud Cold Hardiness and Cultural Safety Zone in Rabbiteye Blueberry Cultivars (래빗아이 블루베리 품종별 꽃눈의 내동성과 재배안전지역)

  • Kim, Hong-Lim;Kwack, Yong-Bum;Han, Jeom-Hwa;Oh, Pill-Kyung;Chae, Won-Byoung;Kim, Seong-Cheol;Kim, Mok-Jong;Kim, Jin-Gook
    • Korean Journal of Environmental Agriculture
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    • v.33 no.4
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    • pp.327-333
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    • 2014
  • BACKGROUND: Rabbiteye blueberry(Vaccinium ashei) is one of the most widely grown blueberry types in the world, together with Northern and Southern highbush blueberry(Vaccinium corymbosum). Rabbiteye blueberry have higher soil adaptability and fruit productivity but less cold tolerance to low temperature than highbush blueberry. The objective of this study is to investigate freezing tolerance of floral buds and establish a cultivation zone for rabbiteye blueberry cultivars. METHODS AND RESULTS: Bearing branches which have similar thickness and same number of floral buds were collected in the early January at the blueberry germplasm preservation plot located in Namhae Sub-station, National Institute of Horticultural and Herbal Science in Gyeongsangnamdo. Cold response of bearing branches were investigated by electrolyte leakage and freezing tolerance of floral buds were determined by ovary browning ratio of 50%($LT_{50}$). Cultivation zone was established based on mean annual extreme minimum temperature for 30 years, from 1981 to 2010. The electrolyte leakage of bearing branches in rabbiteye blueberry increased as temperature decreased and was lowest in 'Brightwell' but highest in 'Bluegem' when they were kept in $-5^{\circ}C$. Besides, the electrolyte leakage increased in 'Brightblue', 'Brightwell', 'Climax', 'Delite', 'Gardenblue', 'Southland' and 'Woodard' in $-20^{\circ}C$. Freezing tolerance($LT_{50}$) was lowest in 'Bluegem' and 'Homebell'($-13.3^{\circ}C$), and highest in 'Tifblue'($-25^{\circ}C$) among different rabbiteye blueberry cultivars. $LT_{50}$ of 'Southland' was from -15.0 to $-16.7^{\circ}C$, that of 'Delite', 'Brightwell',' Austin' and 'Climax' was $-18.3^{\circ}C$, and that of 'Bluebelle', 'Woodard' and 'Powderblue' was $-20^{\circ}C$. CONCLUSION: This study indicate that The hardiness zones of rabbiteye blueberry were classified into Six cultivation zones and cultivation zones of most cultivars were the south of Jeollanam-do and Gyeongdangnam-do, except for 'Tifblue.'