복숭아 네 품종 ‘오도로끼’, ‘가납암백도’, ‘진미’, ‘장호원황 도’ 신초의 저온순화 및 탈순화 동안 시기별 내한성 변화는 전해질 누출률을 분석하여 나타냈다. 또한 내한성 결정 요인 을 분석하고자 SDS-PAGE를 이용하여 dehydrin 함량 변화를 확인하였으며, 그와 관련된 유전자 발현 분석은 quantitative real-time RT-PCR을 이용하여 수행하였다. 네 품종의 내한성 은 2012년 12월까지 꾸준히 증가하였으며 그 후 2013년 4월 까지 감소하였다. PpDhn1 유전자가 인코딩하는 60kDa의 dehydrin 단백질은 탈순화기(2013년 3-4월)에 비하여 저온순화 기(2012년11월-2013년 1월) 동안 높은 축적이 확인되었다. PpDhn1 유전자와 PpDhn3 유전자 발현양상은 복숭아 네 품 종에서 내한성 변화와 평행하게 나타난 반면, PpDhn2 유전자 는 뚜렷한 시기별 패턴을 나타내지 않았다.
We investigated Arctic plants to determine if they have a specific mechanism enabling them to adapt to extreme environments because they are subject to such conditions throughout their life cycles. Among the cell defense systems of the Arctic mouse-ear chickweed Cerastium arcticum, we identified a stress-responsive dehydrin gene CaDHN that belongs to the SK5 subclass and contains conserved regions with 1 S-segment at the N-terminus and 5 K-segments from the N-terminus to the C-terminus. To investigate the molecular properties of CaDHN, yeast were transformed with CaDHN. CaDHN-expressing transgenic yeast (TG) cells recovered more rapidly from challenge with exogenous stimuli, including oxidants (hydrogen peroxide, menadione, and tert-butyl hydroperoxide), high salinity, freezing and thawing, and metal (Zn2+), than wild-type (WT) cells. TG cells were sensitive to copper, cobalt, and sodium dodecyl sulfate. In addition, the cell survival of TG cells was higher than that of WT cells when cells at the mid-log and stationary stages were exposed to increased ethanol concentrations. There was a significant difference in cultures that have an ethanol content >16%. During glucose-based batch fermentation at generally used (30℃) and low (18℃) temperatures, TG cells produced a higher alcohol concentration through improved cell survival. Specifically, the final alcohol concentrations were 13.3% and 13.2% in TG cells during fermentation at 30℃ and 18℃, respectively, whereas they were 10.2% and 9.4%, respectively, in WT cells under the same fermentation conditions. An in vitro assay revealed that purified CaDHN acted as a reactive oxygen species (ROS)-scavenger by neutralizing H2O2 and a chaperone by preventing high temperature-mediated catalase inactivation. Taken together, our results show that CaDHN expression in transgenic yeast confers tolerance to various abiotic stresses by improving redox homeostasis and enhances fermentation capacity, especially at low temperatures (18℃).
더덕 뿌리에서 유래한 EST 라이브러리로부터 dehydrin 유전자와 높은 상동성 을 나타내는 full clone cDNA를 얻었다. 더덕의 dehydrin, ClDhn1은 893 bp의 cDNA로 159개의 아미노산을 코딩하는 480 bp의 ORF를 가지고 있다. ClDhn1의 아미노산을 분석해 보면, 전체적으로 높은 친수성을 나타내며, lysine이 풍부한 K 반복구간(KIKEKLPG)을 카르보닐기 쪽에 2개 가지고 있다. 또한, 여러 dehydrin들의 공통적인 특징인 7개의 연속적인 serine잔기가 첫 번째 K반복 구간 앞에 위치한다. 그러나, 아미노기쪽의 DEYGNP보존 구간은 변형(DEHGNP)되어 있다. ClDhn1 유전자는 전사 단계에서 더덕의 뿌리에서 가장 높은 발현 양상을 보이며, 줄기와 잎에서는 적은 양이 발현되었다.
Dehydrins (LEA Dll proteins) are one of the typical families of plant proteins that accumulate in response to dehydration, cold stress, abscisic acid, or during seed maturation. A 1.3-kb cDNA was cloned from a cDNA expression library of 5-day-old germinating maize scutellums under drought stress. The deduced protein sequence indicated a dehydrin gene encoding SK3 LEA protein typically expressed during cold acclimation, but not by drought stress in barley and wheat. Thus, it was named maize DEHYDRIN2 (ZmDhn2). It accumulates rapidly and highly in drought-stressed scutellum and leaf tissues at any stage, but not under cold stress. ZmDhn2 gene was transformed into Arabidopsis thaliana for functional analysis under drought condition. From electrolyte leakage test, no significant difference showed between wild type and transformants under normal growth condition, but the leakage level of electrolyte in wild type plants was about 3 times as high as that in the transformed plants under drought stress. It suggests that ZmDHN2 playa role in increasing drought tolerance.