Sweetpotato (Ipomoea batatas L.) is a globally important food crop that is susceptible to infestation with the root-knot nematode, Meloidogyne incognita, which causes substantial crop losses. Previous transcriptomic and proteomic analyses identified several genes that displayed differential expression patterns in susceptible and resistant cultivars in response to root-knot nematode (RKN) infection. As a result of previous study, RKN infection was confirmed in the RKN-susceptible sweetpotato cultivar Yulmi. Transcriptome analysis confirmed that among the genes that respond in this process, there are many genes related to ethylene biosynthesis. Therefore, in this study, we focused on the ACC oxidase (ACO) gene, the final enzyme of ethylene biosynthesis, and analyzed the expression patterns under various abiotic stress conditions. Using transcriptome data from our previous study, various expression changes in the four ACO genes used in this study were confirmed during RKN infection. The expression of G25011|TU41034 decreased during RKN infection compared to the untreated control, while the expression of G31097|TU51009, G28360|TU46486, and G15447|TU25395 genes increased in the early stages of RKN infection. Expressions of four ACO genes in leaves of sweetpoato were investigated under abiotic stress conditions such as wounding, high salinity, dehydration, and low temperature stress treatment. Expression of the G25011|TU41034 was significantly increased under abiotic stress conditions except low temperature. G31097|TU51009 was hardly expressed under abiotic stress conditions. Although the expression pattern of G28360|TU46486 and G15447|TU25395 was slightly different depending on the type of abiotic stress, an overall increase in expression was observed. It is expected that this study will be used as basic data on how ethylene biosynthesis responds not only to nematode infection but also to various abiotic stress conditions and will be helpful in functional studies of various crops.
Forage crop management is severely challenged by global warming-induced climate changes representing diverse a/biotic stresses. Thus, screening of valuable genetic resources would be applied to develop stress-tolerant forage crops. We isolated two NAC (NAM, ATAF1, ATAF2, CUC2) transcription factors (ANAC032 and ANAC083) transcriptionally activated by multi-abiotic stresses (salt, drought, and cold stresses) from Arabidopsis by microarray analysis. The NAC family is one of the most prominent transcription factor families in plants and functions in various biological processes. The enhanced expressions of two ANACs by multi-abiotic stresses were validated by quantitative RT-PCR analysis. We also confirmed that both ANACs were localized in the nucleus, suggesting that ANAC032 and ANAC083 act as transcription factors to regulate the expression of downstream target genes. Promoter activities of ANAC032 and ANAC083 through histochemical GUS staining again suggested that various abiotic stresses strongly drive both ANACs expressions. Our data suggest that ANAC032 and ANAC083 would be valuable genetic candidates for breeding multi-abiotic stress-tolerant forage crops via the genetic modification of a single gene.
Cold, salt and heat are the most critical factors that restrict full genetic potential, growth and development of crops globally. However, clarification of genes expression and regulation is a fundamental approach to understanding the adaptive response of plants under unfavorable environments. In this study, we applied an annealing control primer (ACP) based on the GeneFishing approach to identify differentially expressed genes (DEGs) in Italian ryegrass (cv. Kowinearly) leaves under cold, salt and heat stresses. Two-week-old seedlings were exposed to cold (4°C), salt (NaCl 200 mM) and heat (42°C) treatments for six hours. A total 8 differentially expressed genes were isolated from ryegrass leaves. These genes were sequenced then identified and validated using the National Center for Biotechnology Information (NCBI) database. We identified several promising genes encoding light harvesting chlorophyll a/b binding protein, alpha-glactosidase b, chromosome 3B, elongation factor 1-alpha, FLbaf106f03, Lolium multiflorum plastid, complete genome, translation initiation factor SUI1, and glyceraldehyde-3-phosphate dehydrogenase. These genes were potentially involved in photosynthesis, plant development, protein synthesis and abiotic stress tolerance in plants. However, this study provides new insight regarding molecular information about several genes in response to multiple abiotic stresses. Additionally, these genes may be useful for enhancement of abiotic stress tolerance in fodder crops as well a crop improvement under unfavorable environmental conditions.
Glutathione S-transferase (GST) is a key gene involved in multiple stress tolerance in all living organisms, though it is still to be disclosed the gene function in teff grass [Eragrostis tef (Zucc.)Trotter].The objectives of this study were to clone and molecular characterization of GST gene in teff grass. We characterized GST1 from teff grass (EtGST1), it composed of a 645-bp open reading frame (ORF) that encoded 195 amino acid residue. Further, we transformed EtGST1 in E.coli BL21 (DE3) cells. This recombinant EtGST1 in E.coli BL21(DE3) induced at 37°C temperature. In addition, Growth of cells overexpressing EtGST1 rapidly increased in the presence of polyethylene glycol (5%), heat (46°C), NaCl (0.6%), and arsenic (1 mM) than that of cells harboring an empty vector. These results suggest that EtGST1 would be suitable candidate for improving tolerance in forages and/or grasses species against multiple abiotic stresses.
Drought is one of the detrimental factors that impair plant growth and productivity. In this study, we applied annealing control primer (ACP)-based reverse transcriptase PCR (polymerase chain reaction) technique to identify differentially expressed genes (DEGs) in maize leaves in response to drought stress. Two-week-old maize seedlings were exposed to drought (DT) by suspending water supply. DEGs were screened after 3 days of DT-treated samples using the ACP-based technique. Several DEGs encoding 16.9 protein, antimicrobial protein, hypothetical protein NCLIV_068840, thioredoxin M-type were identified in maize leaves under drought stress. These genes have putative functions in plant defense response, growth and development. These identified genes would be useful for predictive markers of plant defense, and growth responses under drought stress in plants.
규산은 작물의 필수원소에는 포함되어있지 않으나, 화본과 작물을 중심으로 내도복성과 병충해 저항성의 향상 , 군락구조 개선에 의한 광합성 능력의 향상 등에서 폭넓게 그 유용성이 알려져 왔으며, 최근에는 원예작물에서도 규산질 비료의 시용이 수량이나 병충해저항성을 향상시키는 효과가 입증되고 있어 친환경농업 관점에서도 주목을 밭고 있다. 본 실험은 배추 육묘 중 규산질 비료의 시용이 묘소질과 저온, 고온, 건조 등 환경내성에 미치는 영향을 검토하기 위하여 수행하였다. 규산염 처리농도를 8, 16, 32, 64 및 128mM로 설계 하여 주 2회 관주 처리 하고, 처리 3주 후에 생육조사 및 스트레스 내성에 대해 평가하였다. 생육조사 결과, 8, 16 및 32m의 농도에서는 대부분의 생육지표가 대조구에 비해 약간 증가하는 경향을 보였으나 8mM처리만 제외하고 통계적 유의차는 나타나지 않았다. 고농도인 128mM의 규산 처리구에서는 모든 생육 지표가 감소하였다. 총 뿌리 면적, 뿌리 길이 및 근단 수는 8, 16 및 32mM의 농도에서 증가했지만 64 및 128mM의 처리구 에서는 감소하였다. 규산 처리 농도가 증가함에 따라 증산 속도는 감소한 반면 기공확산 저항은 증가하는 경향을 보였다. 상대적 이온 누출율도 대조구에 비해 규산염 처리구에서 감소되었으나, 처리 농도간 유의차는 나타나지 않았다. 규산처리에 의해 고온과 저온 장해 지표도 감소되었으며, 농도간에는16과 32mM이 가장 효과적이었다. 규산처리에 따라 건조내성도 증가하여 대조구는 단수 후 3일째부터 위조되기 시작하여 5일째는 전 개체가 위조하였으나, 규산처리구는 4일(8, 64, 128 mM) 또는 5일(16과 32mM) 부터 위조가 시작되어 6일 (8mM)이나 7일(16, 32 ,64및 128 mM)이 지나서야 모든 공시 개체가 위조되었다.
본 연구는 환경스트레스 저항성이 증진된 페튜니아를 개발하기 위하여 NDPK2유전자 도입 형질전환 계통 NDPK2-7-1와 SOD2 유전자 도입 형질전환 계통 SOD2- 2-1-1-35간의 교잡에 의해 획득된 후대들의 비생물적 스트레스 저항성을 조사하기 위해 수행되었다. 비 생물적 스트레스 유발원인 메틸바이올로젠(methyl viologen, MV) 100 μM과 200 μM 처리에서 교잡후대들은 그들의 교배 모본 SOD2 유전자나 NDPK2 유전자가 단독으로 도 입된 형질전환 계통이나 비형질전환체 보다 메틸바이 올로젠에 의한 피해를 적게 받았다. 이는 SOD2 유전 자나 NDPK2 유전자가 단독으로 도입된 형질전환 계 통간 교잡에 의해 획득된 후대들이 그들의 교배모본 (SOD2 유전자나 NDPK2 유전자가 단독으로 도입된 형질전환 계통)이나 비형질전환체 보다 산화적 스트레 스에 대한 저항성이 증진되었음을 증명해 준다고 할 수 있다. 이들 교잡후대들은 초장 등 11종류의 양적형질의 특성이 비형질전환체에 비해 약간 길거나 짧긴 하였지 만 비형질전환체와 거의 유사하였으며, 꽃 색갈이나 모양 또한 그들의 교배모본 (SOD2 유전자나 NDPK2 유전 자가 단독으로 도입된 형질전환 계통)이나 비형질전환 체와 차이가 없었다.
In the present study, we have used an annealing-control-primer (ACP)-based differentially display RT-PCR method to identify salt-stress-induced differentially expressed genes (DEGs) in barley leaves. Using 120 ACPs, a total of 11 up-regulated genes were identified and sequenced. Temporal expression patterns of some up-regulated DEGs in response to salt stress were further analyzed by Northern blot analysis. The possible roles of these identified genes are discussed within the context of their putative role in response to salt stress. Thus, the identification of some novel genes-such as SnRK1-type protein kinase; 17 kDa, class I, small heat shock protein; and RNase S-like protein precursor genes-may offer a new avenue for better understanding the salt stress response in plants, knowledge which might be helpful for developing future strategies.
ABA는 식물에서 비 생물학적 스트레스 내성에 관여하는 중 요한 식물 호르몬이다. 애기장대의 group A bZIP 전사인자는 ABA 신호전달 과정에 중요한 역할을 한다고 알려져 있다. 그러나 벼에서는 group A bZIP 전사인자의 기능이 잘 알려져 있지 않다. 따라서 우리는 벼에서 group A bZIP 전사인자인 OsABF3 (Oryza sativa ABA responsive element binding factor 3)를 연 구하였다. 이를 위해 벼의 다양한 조직과 다양한 스트레스(가 뭄, 염분, 저온, ABA, 산화 스트레스)에 따른 OsABF3 발현 패턴을 분석하였다. 또한 maize의 원형질체에서 GFP fusion 벡터를 사용한 세포 내 위치 분석을 통해 OsABF3가 핵단백질이라는 것을 확인하였다. Yeast one-hybrid 실험을 통해 OsABF3의 Cterminal 부분이 ABREs에 결합한다는 것과 N-terminal 부분 이 하위 유전자의 transactivation 하는데 필요하다는 것을 알수 있었다. 그리고 T-DNA가 삽입된 OsABF3의 homozygous 돌연변이체가 야생형과 과발현체에 비해 발아와 발아 후 단계 에서 고농도의 ABA에 대한 민감도가 더 감소한 것을 알 수 있었다. 결과적으로 종합해 볼 때 OsABF3는 ABA의 의존적인 경로를 통해 비 생물학적 스트레스에 반응하는 유전자의 발현을 조절하는 기능을 하는 전사 조절자이다. 또한 OsABF3의 transactivation을 측정하는 실험에 있어서 억제 domain이 존 재한다는 결과를 얻었다.
Background : Panax ginseng C. A. Meyer is a slow-growing perennial herb that is cultivated in shading condition. Climate change occur around the world that make a lot of problem such as damage of high temperature, drought, salinity and disease. The problems lower the ginseng productivity that cause income reduction of farmers. To achieve stable ginseng production, development of elite varieities resistant to abiotic and biotic stresses is consistently required. It is very time consuming process in order to develop new ginseng varieties because ginseng flowers after 3 years of growth. So, early selection system of elite line must be established. This study was conducted to develope efficient ginseng breeding techniques for early identification of heat or salinity resistance. Methods and Results : Ginseng petioles was soaked in mixed salts solution consisting of KNO3, KH2PO4, MgSO4․H2O in order to test resistant or susceptible salinity. The degree of resistance was quantified according to damage size. Also, ginseng lines transplanted in pot were treated 46℃ for 1 hour and then chlorophyll fluorescence reaction were measured in order to test resistant or susceptible high-temperature. The measured values such as Fm/Fo, Fv/Fm, Rfd were differentiated between resistant and susceptible line. Conclusion : Several lines showed that they are resistance to high temperature or salinity. The selected lines will be utilized for parents to develop new varieties.
UDP-glucose 4-epimerase (UGE; EC 5.1.3.2) is an enzyme that plays an essential role in the interconverts UDP-D-glucose (UDP-Glc) and UDP-Dgalactose (UDP-Gal). Five members of the Chinese cabbage (Brassica rapa) UDP-glucose 4-epimerase gene family, designated BrUGE1 to BrUGE5, have been cloned and characterized. Quantitative PCR shows that the BrUGE1and BrUGE4 mRNA are most abundant among other BrUGE genes, accounting for more than 55% of total BrUGE transcripts in most of the tissues examined. All genes showed organ specific expression pattern, two of which (BrUGE1 and 4) actively responded after Pectobacterium carotovorum subsp. carotovorum infection, while four genes (BrUGE-1, -3, -4 and -5)were shown to respond considerably against salt, drought and abscisic acid (ABA) treatments. To better understand the function of the UGE gene, we constructed a recombinant pART vector carrying the BrUGE1 gene under the control of the CaMV 35S promoter and nos terminator and transformed using Agrobacterium tumefaciens. We then investigated BrUGE1 overexpressing rice lines at the physiological and molecular levels under biotic and abiotic stress conditions. Bioassay of T3 progeny lines of the transgenic plants in Yoshida solution containing 120 mM Nacl for 2 weeks, confirmed that the BrUGE1 enhances salt tolerance to transgenic rice plants. Also T3 progeny lines of the transgenic plants, when exposed to infection caused by Xanthomonas oryzae pv oryzae, showed tolerance to bacterial blight. These results showed that BrUGE1 can be used as potential genetic resource for engineering Brassica with multiple stress resistance.
The plant-specific NAC (NAM, ATAF, and CUC)-domain proteins play important roles in plant development and stress responses. Comparative time-course expression analyses were carried out to analyze the expression levels of 62 soybean NAC genes during drought stress in order to search for the stress-inducible NAC genes. Ten GmSNAC (Glycine max stress-inducible NAC) genes having the significant differential expression in response to the drought stress and abscisic acid (ABA) hormone application were further investigated for their expression profiles with various stresses such as drought, high salinity, cold and with ABA treatments by the quantitative real-time PCR analyses. In this research, the full-length cDNAs of eight GmSNAC were isolated for the further studies. Eight GmSNAC proteins were tested for their transcription activation in the yeast assay system. Two GmSNAC proteins showed the very high transcriptional activities and the other two GmSNAC proteins displayed moderate levels of transactivation while the remaining four GmSNAC proteins lacked transactivation in yeast. Subcellular localization of eight GmSNAC proteins was analyzed via the green fluorescent protein-GmSNAC fusion protein in tobacco plant cell. Three GmSNAC proteins with the C-terminal transmembrane domain were localized to the nucleus and cytoplasmic fractions. The other five GmSNAC proteins were targeted to the nucleus. The function of GmSNAC49 gene was further investigated using the overexpression transgenic Arabidopsis. Germination rate in transgenic plants over-expressing GmSNAC49 was delayed in the media supplemented with mannitol or ABA compared with that of wild-type (WT) plants. The 35S:GmSNAC49 transgenic Arabidopsis displayed improved tolerance to drought stress compared to the WT. The results of this systematic analysis of the GmSNAC family responsive to abiotic stress will provide novel tools and resources for the development of improved drought tolerant transgenic soybean cultivars