• Title/Summary/Keyword: Abiotic Stress

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Construction and Analysis of Binary Vectors for Co-Overexpression, Tissue- or Development-Specific Expression and Stress-Inducible Expression in Plant (식물에서 표적 유전자의 동시 과발현, 조직/발달 특이적 발현 및 스트레스 유도성 발현을 위한 binary 벡터의 제작과 분석)

  • Lee, Young-Mi;Park, Hee-Yeon;Woo, Dong-Hyuk;Seok, Hye-Yeon;Lee, Sun-Young;Moon, Yong-Hwan
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1314-1323
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    • 2010
  • In this study, we constructed various kinds of binary vectors with the pPZP backbone for co-overexpression, tissue- or development-specific expression and stress-inducible expression, and validated them for ectopic expression of target genes. Using a modified CaMV 35S promoter, a binary vector was generated for co-overexpression of two different genes and was confirmed to be efficient for overexpressing two different target genes at the same time and place. Binary vectors containing At2S3, KNAT1 or LFY promoters were constructed for tissue-specific or development-specific gene expression, and the binary vectors were suited for embryo/young seedling stage-, shoot apical meristem- or leaf primordia-specific expressions. Furthermore, the binary vectors containing RD29A or AtNCED3 promoters were validated as suitable vectors for gene expression induced by abiotic stresses such as high salt, ABA, MV and low temperature. Taken together, the binary vectors constructed in this study would be very useful for analyzing the biological functions of target genes and molecular mechanisms through ectopic expression.

Overexpression of NtHSP70-1 Protects Chlorophyll from High Temperature in Plants (NtHSP70-1에 의한 클로로필의 고온 내성 효과)

  • Cho, Eun-Kyung;Hong, Choo-Bong
    • Journal of Life Science
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    • v.18 no.3
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    • pp.304-310
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    • 2008
  • Heat shock protein 70 (HSP70) is known as molecular chaperone, the fundamental protein participating in various processes, from nascent protein synthesis to protection of proteins during abiotic stresses and developmental programs. However, their biological functions in plants are not yet well known. Here, NtHSP70-1 (AY372069), HSP70 of Nicotiana tabacum induced by heat stress was investigated. To analyze the protective role of NtHSP70-1, transgenic tobacco plants, which constitutively overexpressed NtHSP70-1 as well as contained either the vector alone or having NtHSP70-1 in the antisense orientation, were constructed. The altered NtHSP70-1 levels in plants were confirmed by western blotting and transgenic sense lines exhibited tolerance to heat stress. Seedlings with the constitutively expressed NtHSP70-1 grew as green or healthy plants after heat stress. In contrast, transgenic vector or antisense lines exhibited yellowing of leaves or some delay in growth, which finally led to death. Evaluation of chlorophyll contents of heat-shocked transgenic tobacco seedlings indicated that NtHSP70-1 contributes to thermotolerance by preventing chlorophyll synthesis in plants.

Isolation and characterization of a monodehydroascorbate reductase gene in poplar (Populus alba × P. glandulosa) (현사시나무 monodehydroascorbate reductase (MDHAR) 유전자의 분리 및 발현특성)

  • Yoon, Seo-Kyung;Park, Eung-Jun;Bae, Eun-Kyung;Choi, Young-Im;Kim, Joon-Hyeok;Lee, Hyoshin
    • Journal of Plant Biotechnology
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    • v.41 no.4
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    • pp.194-200
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    • 2014
  • Monodehydroascorbate reductase (MDHAR) is an important enzyme that plays a role in the detoxification of reactive oxygen species (ROS) by maintaining reduced pool of ascorbate through recycling the oxidized form of ascorbate. In this study, we isolated a PagMDHAR1 gene from Populus alba ${\times}$ P. glandulosa, and investigated its expression characteristics. The PagMDHAR1 cDNA encodes a putative 434 amino acids containing FAD- and NAD(P)H-binding domains. Southern blot analysis indicated that a single nuclear gene encodes this enzyme. Northern hybridization analysis revealed that PagMDHAR1 is highly expressed in both suspension cells and flower tissues, while its expression levels were enhanced by drought, salt, cold, wounding and ABA. Therefore, PagMDHAR1 might be expressed in response to abiotic stress through the ABA-mediated signaling pathway in this poplar species, suggesting that the PagMDHAR1 plays an important role in the defense mechanisms against oxidative stress.

Effects of Short Microwave Irradiation Time at the Seedlings Stage on the Growth and Secondary Metabolite Contents of Lettuce (Lactuca sativa L.) (유묘단계에서 단시간 마이크로웨이브 처리가 상추의 생육 및 이차대사산물 함량에 미치는 영향)

  • Yong Jae Lee;Su Yong Park;Ju Hyung Shin;Seung Yong Hahm;Gwang Ya Lee;Jong Seok Park
    • Journal of Bio-Environment Control
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    • v.32 no.3
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    • pp.217-225
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    • 2023
  • This experiment was conducted to investigate the effects of microwave irradiation on the growth and secondary metabolite contents of lettuce seedlings. Seedlings at three weeks after sowing were treated by a microwave oven for 0, 4, 8, and 12 seconds with 200 W. After cultivation in a close plant production system for 4 weeks, plant growth measurements and secondary metabolite analysis were performed. The results showed that the fresh and dry weights of the shoot and root, leaf area, leaf length, and the number of leaves were decreased as increasing the microwave treatment times. Chlorophyll a and b, total carotenoids were increased and total phenolics were decreased at the 12-second treatment compared to the 4-second treatment. Total flavonoid contents were decreased at the 8-second treatment compared to the control. These results suggest that the changes in the levels of secondary metabolites were caused by oxidative stress. Although there was no significant difference in secondary metabolite contents excluding total flavonoid contents on the microwave treatments compared to the control, the significant difference suggests that the microwave treatment of 200 W and 2.45 GHz may alter secondary metabolite contents of lettuce after 4 weeks.

Expression of CsRCI2s by NaCl stress reduces water and sodium ion permeation through CsPIP2;1 in Camelina sativa L.

  • Kim, Hyun-Sung;Lim, Hyun-Gyu;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.194-194
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    • 2017
  • Camelina (Camelina sativa L.) is a potential bio-energy crop that has short life cycle about 90 days and contains high amount of unsaturated fatty acid which is adequate to bio-diesel production. Enhancing environmental stress tolerance is a main issue to increase not only crop productivity but also big mass production. CsRCI2s (Rare Cold Inducible 2) are cold and salt stress related protein that localized at plasma membrane (PM) and assume to be membrane potential regulation factor. These proteins can be divide into C-terminal tail (CsRCI2D/E/F/G) or no-tail group (CsRCI2A/B/C/H). However, function of CsRCI2s are less understood. In this study, physiological responses and functional characterization of CsRCI2s of Camelina under salt stress were analyzed. Full-length CsRCI2s (A/B/E/F) and CsPIP2;1 sequences were confirmed from Camelina genome browser. Physiological investigations were carried out using one- or four-week-old Camelina under NaCl stress with dose and time dependent manner. Transcriptional changes of CsRCI2A/B/E/F and CsPIP2;1 were determined using qRT-PCR in one-week-old Camelina seedlings treated with NaCl. Translational changes of CsRCI2E and CsPIP2;1 were confirmed with western-blot using the antibodies. Water transport activity and membrane potential measurement were observed by cRNA injected Xenopus laevis oocyte. As results, root growth rate and physiological parameters such as stomatal conductance, chlorophyll fluorescence, and electrolyte leakage showed significant inhibition in 100 and 150 mM NaCl. Transcriptional level of CsPIP2;1 did not changed but CsRCI2s were significantly increased by NaCl concentration, however, no-tail type CsRCI2A and CsRCI2B increased earlier than tail type CsRCI2E and CsRCI2F. Translational changes of CsPIP2;1 was constitutively maintained under NaCl stress. But, accumulation of CsRCI2E significantly increased by NaCl stress. CsPIP2;1 and CsRCI2A/B/E/F co-expressed Xenopus laevis oocyte showed decreased water transport activity as 61.84, 60.30, 62.91 and 76.51 % at CsRCI2A, CsRCI2B, CsRCI2E and CsRCI2F co-expression when compare with single expression of CsPIP2;1, respectively. Moreover, oocyte membrane potential was significantly hyperpolarized by co-expression of CsRCI2s. However, higher hyperpolarized level was observed in tail-type CsRCI2E and CsRCI2F than others, especially, CsRCI2E showed highest level. It means transport of $Na^+$ ion into cell is negatively regulated by expression of CsRCI2s, and, function of C-terminal tail is might be related with $Na^+$ ion influx. In conclusion, accumulation of NaCl-induced CsRCI2 proteins are related with $Na^+$ ion exclusion and prevent water loss by CsPIP2;1 under NaCl stress.

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Effect of Exogenous Application of Salicylic Acid or Nitric Oxide on Chilling Tolerance and Disease Resistant in Pepper Seedlings (외생 살리실산과 일산화질소 처리가 고추묘의 저온 내성 및 병 저항성에 미치는 영향)

  • Park, Song-Yi;Kim, Heung-Tae;Oh, Myung-Min
    • Journal of Bio-Environment Control
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    • v.23 no.4
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    • pp.329-336
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    • 2014
  • As an abiotic stress, chilling stress is one of the major factors limiting plant growth and increasing susceptibility to pathogens. Therefore, enhancing stress tolerance in plants is an important strategy for their survival under unfavorable environmental conditions. The objective of this study was to determine the effects of the exogenous application of salicylic acid (SA) or nitric oxide (NO) on chilling tolerance in pepper seedlings. Pepper (Capsicum annuum L. 'kidaemanbal') seedlings were grown under normal growing conditions ($20/25^{\circ}C$, 15 hours photoperiod, $145{\pm}5{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, fluorescence lamps) for 23 days after transplanting. The solution (3 mL) of 1 mM SA and 0.3 mM NO with surfactant triton 0.1% were sprayed two times a week, respectively. Right after the completion of chemical application, seedlings were subjected to chilling condition at $4^{\circ}C$ for 6 hours under dark condition and then the seedlings were recovered at the normal growing conditions for 2 days. In order to assess plant tolerance against chilling stress, growth characteristics, chlorophyll fluorescence (Fv/Fm), and membrane permeability were determined after chilling stress imposition. Total phenolic concentration and antioxidant capacity were measured during the whole experimental period. Disease incidence for pepper bacterial spot and wilt was also analyzed. Pepper seedlings treated with SA or NO were maintained similar dry mass ratio, while the value in control increased caused by chilling stress suggesting relatively more water loss in control plants. Electrolyte leakage of pepper seedlings treated with SA or NO was lower than that of control 2 days after chilling treatment. Fv/Fm rapidly decreased after chilling stress in control while the value of SA or NO was maintained about 0.8. SA increased higher total phenolic concentration and antioxidant capacity than NO and control during chemical treatment. In addition, increase in total phenolic concentration was observed after chilling stress in control and NO treatment. SA had an effect on the reduction of bacterial wilt in pepper seedlings. The results from this study revealed that pre-treatment with SA or NO using foliar spray was effective in chilling tolerance and the reduction of disease incidence in pepper seedlings.

Effects of Long-Term Subcultured Arbuscular Mycorrhizal Fungi on Red Pepper Plant Growth and Soil Glomalin Content

  • Selvakumar, Gopal;Yi, Pyoung Ho;Lee, Seong Eun;Shagol, Charlotte C.;Han, Seung Gab;Sa, Tongmin;Chung, Bong Nam
    • Mycobiology
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    • v.46 no.2
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    • pp.122-128
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    • 2018
  • Arbuscular mycorrhizal fungi (AMF) are well-known for their ability to improve plant growth and help plants withstand abiotic stress conditions. Unlike other fungi and bacteria, AMF cannot be stored, as they are obligate biotrophs. Long-term preservation of AMF spores is challenging and may lead to the loss of viability and efficiency. This study aimed to understand the effect of prolonged subculture of AMF species on the growth and glomalin-related soil protein (GRSP) from red pepper (Capsicum annuum L.). AMF spores were mass-produced using different techniques and subcultured in pots with sorghum sudangrass as the host plant for 3 years. Experimental soil samples were collected from natural grassland. Five different AMF inocula were used in triplicate as treatments. After 70 days of growth, red pepper plants were harvested and plant dry weight, plant nutrient content, mycorrhizal colonization, AMF spore count, and soil glomalin content were determined. AMF-treated plants displayed higher dry weight than controls, with only fruit dry weight being significantly different. Similarly, significant differences in phosphorous and potassium contents of the above-ground plant parts were observed between mycorrhizal and control treatments. In addition, soil GRSP content was significantly higher in plants inoculated with Rhizophagus sp. and Gigaspora margarita. The increased plant growth and GRSP content suggest that AMF can be maintained for 3 years without losing their efficiency if subcultured regularly with different symbiotic host plants.

Overexpression of Gene Encoding Tonoplast Intrinsic Aquaporin Promotes Urea Transport in Arabidopsis

  • Kim, Sun-Hee;Kim, Kang-Il;Ju, Hyun-Woo;Lee, Ho-Joung;Hong, Suk-Whan
    • Journal of Applied Biological Chemistry
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    • v.51 no.3
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    • pp.102-110
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    • 2008
  • Complementation assay of the urea uptake-defective yeast mutants led to the identification of the Arabidopsis AtTIP4;1 gene encoding the aquaporin. However, its physiological functions still remain elusive. In the present study, histochemical and genetic analyses were performed to understand the physiological roles of AtTIP4;1 in urea uptake. The AtTIP4;1 product was detectible in the roots, but not in the leaves, the stem, and the flower. Its promoter allowed the expression of the $\beta$-glucuronidase reporter gene in the roots and the apical meristem in Arabidopsis. The AtTIP4;1 products were induced under nitrogen-deficient conditions. To investigate the role of the tonoplast intrinsic protein in urea transport and developments, Arabidopsis with the loss- and the gain-of-function mutations by T-DNA insertion in AtTIP4;1 and 35S promoter-mediated overexpression of AtTIP4;1 were identified, respectively. The transfer DNA insertion and the AtTIP4;1-overexpressed plants showed normal growth and development under normal or abiotic stress growth conditions. The urea-uptake studies using $^{14}C$-labeled urea revealed higher accumulation of urea in the AtTIP4;1-overexpressed plants. These results provide evidence that overexpression of AtTIP4;1 leads to the increase in the urea-uptake rate in plants without detectable defects to the growth and development.

Selection of Tolerant Plant Species using Pot Culture for Remediation of Explosive Compounds Contaminated Soil (포트 재배에 의한 화약물질 오염토양 정화용 내오염성 식물 선정)

  • Lee, Ahreum;Bae, Bumhan
    • Journal of Soil and Groundwater Environment
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    • v.20 no.6
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    • pp.73-84
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    • 2015
  • Nine plant species were selected through vegetation survey at three military shooting ranges at northern Gyeonggi Province. Plants were germinated in normal soil and three seedlings were transplanted to a bottom sealed pot containing sandy loam soils contaminated with either RDX (291 mg/kg) or TNT (207 mg/kg). Planted, blank (without plant), and control (without explosive compound) pots were grown in triplicate at a green house for 134 days. During cultivation, transplanted plants exhibited chlorosis and necrosis in flower and leaf by explosive toxicity and stress. Only three plants, Wild soybean, Amur silver grass, Reed canary grass, survived in TNT treated pot, while seven plant species except for field penny cress and jimson weed, thrived in RDX treated pot. Appreciable amount of TNT (61.6~241.2 mg/g-D.W.) was detected only in plant roots. Up to 763.3 mg/g-D.W. along with 4-amino-2,6-dinitrotoluene, an intermediate of TNT, accumulated in the root of wild soybean. In addition, azoxy compounds, abiotic intermediates of TNT, were detected in TNT treated soils. RDX absorbed average 1,839.95 mg/kg in shoot and 204.83 mg/kg in root. Most of TNT in plant was accumulated in underground part whereas RDX was localized in aerial part. Material balance calculation showed that more than 95% of the initial TNT was removed in the planted pots whereas only 60% was removed in the blank pot. The amount of RDX removed from soil was in the order of Amur Silver Grass (51%) > Chickweed (43%) > Evening primrose (38%). Based on the results of pot cultures, Amur silver grass and Reed canary grass are selected as tolerant remedial plants for explosive toxicity.

Selection of Kentucky Bluegrass(Poa pratensis L.) Cultivar for Golf Courses in Korea (골프장에 적합한 켄터키 블루그래스 품종 선발)

  • Kim, Kyung-Duck;Tae, Hyun-Sook;Kim, Jong-Bo;Jang, Jae-Il;Oh, Sung-Bae
    • Journal of the Korean Institute of Landscape Architecture
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    • v.35 no.1 s.120
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    • pp.88-93
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    • 2007
  • Since the 1990s, the application of Kentucky bluegrass cultivars has increased in newly-constructed Korean golf courses as opposed to the previously-used zoysiagrass. However, there have been few reports studying the selection and characterization of these Kentucky bluegrass cultivars under Korean weather conditions. A total of 12 Kentucky bluegrass cultivars were tested for their adaptability in environmental stresses during summer conditions in Korea in order to select the best Kentucky bluegrass cultivars for use in Korean golf courses. This study found that two Kentucky bluegrass cultivars ('Midnight' and 'Bluestone') maintained their green throughout the summer season and had a high degree of root density, as compared to the other cultivars tested. These two cultivars also had a good rate of coverage in the early growth period. These characteristics make them suitable for application on sports fields and golf courses, which receive many divots and the frequent replacement of turfgrass sod caused by both biotic and abiotic stress. Two other cultivars, the 'Ginney' and 'Nuglade', also displayed good visual quality and high rate of coverage under summer conditions. In conclusion, the 'Midnight' and 'Bluestone' cultivars performed well in areas including the maintenance of their green color, the number of roots and the rate of coverage during the summer months. These characteristics are necessary for golf courses sports fields, which receive many divots and requirethe frequent replacement of turfgrass. further research on a range of Kentucky bluegrass cultivars, including a sensitivity test for many pathogens and recovery rates from a variety of stresses, is required in the near future.