• 제목/요약/키워드: Homozygous plant

검색결과 57건 처리시간 0.031초

고추 약배양을 통한 고빈도 배발생 및 식물체 재분화 (High Frequency Embryogenesis and Plant Regeneration in Anther Cultures of Pepper)

  • 안민영;김용권;민성란;정원중;인동수;유장렬
    • 식물조직배양학회지
    • /
    • 제25권5호
    • /
    • pp.319-322
    • /
    • 1998
  • 여러 계통의 고추(Capsicm annuum L.)의 약을 이용하여 식물체 재분화 조건을 확립하였다. 꽃받침 위로 꽃잎이 2mm 올라온 미성숙 화뢰에서 상부로부터 절반 이하로 착색된 약을 채취하여 0.1mg/L 2,4-D와 0.1mg/L kinetin이 첨가된 Dumas 고체배지에 치상하였다. 배양 4주후부터 배가 발생하였으며, 8주후의 배발생 빈도는 최고 58.3%였다. 배는 MS 기본배지로 옮겨주었을 때 95이상이 발아하여 식물체로 재분화되었다.

  • PDF

Transformation and Expression of the PAT Gene in Arabidopsis Tryptophan Mutants

  • Lim, Seon-Hee;Kim, Young-Soon;Cheong, Hyeon-Sook
    • Journal of Plant Biology
    • /
    • 제39권4호
    • /
    • pp.243-247
    • /
    • 1996
  • Phosphoribosylanthranilate transferase (PAT) catalyzes the second step of the tryptophan biosynthetic pathway and is encoded by a single-copy gene that complements all the visible phenotypes of the tryptophan mutant (trp1-100) of Arabidopsis. The trp1-100 is blue fluorescent under UV light becuase it accumulates anthranilate. To obtain a plant with reduced PAT activity, PAT1 genes with several internal deletions in different promoter regions (pHS 101, pHS102, pHS104, pHS105, and pHS107) were induced into trp1-100 via Agrobacterium. Then, homozygous T3 plants were isolated and examined for blue fluorescence. Introduction of the PAT1 gene fusants results in the reversion of fluorescence phenotype except in the case of pHS105. These results prompted us to perform a parallel analysis of anthranilate synthase and PAT interms of the genetic complementation. A plant line carrying pHS105 gene fusant does not completely complement the blue fluorescence but it accumulates less anthranilate than trp1-100. The activity of PAT was reduced in the transgenic mutant as well. The plant carrying these constructs will add to the growing collection of molecular tools for the study of the indolic secondary metabolism.

  • PDF

분자마커를 이용한 토마토 시들음병 race 3 저항성 토마토 유전자원 탐색 (Screening for Resistance to Fusarium oxysporum f. sp. lycopersici Race 3 Using Molecular Marker in Tomato Germplasm)

  • 허온숙;노나영;고호철;김상규;이주희;곽재균;오세종
    • 식물병연구
    • /
    • 제18권4호
    • /
    • pp.304-309
    • /
    • 2012
  • 세 개의 race가 보고되어 있는 Fusarium oxysporum f. sp. lycopersici는 전 세계적으로 그 피해가 막대한 토마토 시들음병의 원인균으로 알려져 있다. 토마토 유전자원 1,906 자원을 대상으로 토마토 시들음병 race 3 저항성 유전자 I-3의 유전형을 high resolution melting 법을 사용하여 분석하였다. 총 97점의 자원에서 homozygous resistant 피크가 확인되었으며, 8점은 이형접합성, 1,801점은 감수성으로 나타났다. 저항성 자원 97점의 종 분포는 S. lycopersicum var. lycopersicum가 65점, S. lycopersicum var. cerasiforme가 13점, 12종의 야생근연종 중에서 저항성 자원은 S. pimpinellifolium 8점, S. habrochaites 3점, S. corneliomulleri 3점, S. galapagense 1점, S. peruvianum 3점, S. chilense 1점으로 총 19점이었다. 하지만 좀 더 정확한 시들음병 저항성 평가를 위하여 추후에 race 1과 2에 대한 유전분석과 생물검정이 필요하다.

염분, 저온 및 가뭄 스트레스 조건에서 벼 ND0001 oscpk11 돌연변이체의 OsCPK11 발현 분석 (Expression Analysis of OsCPK11 by ND0001 oscpk11 Mutants of Oryza sativa L. under Salt, Cold and Drought Stress Conditions)

  • 김현미;김성하
    • 생명과학회지
    • /
    • 제31권2호
    • /
    • pp.115-125
    • /
    • 2021
  • 칼슘-의존성 단백질 카이네이즈(CDPK)는 식물의 Ca2+ 매개 신호 전달에 필수적인 역할을 한다. CDPK는 염분, 저온, 가뭄 등과 같은 비생물적 스트레스에 대한 식물의 반응을 조절하는 데 관여하는 것으로 알려져 있다. 벼의 CDPK는 31개의 유전자로 구성된 거대 다유전자군으로 되어 있지만, 단지 일부 벼의 CDPK 기능만이 확인되었다. 따라서, 벼에서 OsCPK11의 기능을 알아보기 위해, 이 연구는 염분, 저온 및 가뭄과 같은 비생물적 스트레스 조건에서 벼의 야생형과 ND0001 oscpk11 돌연변이체의 OsCPK11 발현 분석에 초점을 맞추었다. 염분, 저온, 가뭄 스트레스 처치를 위해 유식물을 각각 200 mM NaCl, 4℃, 20% PEG 6,000에 노출시켰다. 야생형과 ND0001 돌연변이체에서 OsCPK11의 발현을 확인하기 위해 RT-PCR과 quantitative real-time PCR을 수행하였다. RT-PCR 결과에 의하면, 야생형과 이형접합성 돌연변이체에서는 OsCPK11 전사체가 검출되었지만, 동형접합성 돌연변이체에서는 검출되지 않았다. Quantitative real-time PCR 결과에 의하면 야생형에서 염분, 저온, 가뭄 스트레스에 의해 OsCPK11의 상대적인 발현이 증가하였으며, 각각 24시간, 6시간, 24시간 후 최대 수준에 도달하였다. ND0001 동형접합성 돌연변이체의 OsCPK11의 상대적 발현은 야생형에 비해 현저히 감소하였다. 이러한 결과는 oscpk11 동형접합성 돌연변이체에서는 OsCPK11발현을 완전히 저해하며, OsCPK11유전자 발현 조절이 염분, 저온 및 가뭄 스트레스 신호 전달 과정에 관여할 수 있음을 의미한다.

Absence of AVP1 transcripts in wild type watermelon scions grafted onto transgenic bottle gourd rootstocks

  • Kim, Byung Oh;Han, Jeung-Sul;Park, Kyung Il;Jeon, Su Min;Kim, Chang Kil
    • Journal of Plant Biotechnology
    • /
    • 제42권1호
    • /
    • pp.13-18
    • /
    • 2015
  • In this study we confirmed the stable integration of Arabidopsis AVP1 in the genomes of bottle gourd $T_3$ homozygous lines and its transcription, and additionally evaluated possibility of translocation of the AVP1 mRNA from transgenic bottle gourd rootstocks to wild type watermelon scions. Each AVP1 gene in two bottle gourd T3 lines is abundantly expressed under a field condition. Given the grafting between wild type watermelon scions and AVP1-expressing bottle gourd rootstocks, no translocation of the AVP1 mRNA was detected in leaves, both sexual flowers, and fruits of the scions.

Ac/Ds-mediated gene tagging system in rice

  • Eun, Moo-Young;Yun, Doh-Won;Nam, Min-Hee;Yi, Gi-Hwan;Han, Chang-Deok;Kim, Doh-Hoon;Park, Woong-June;Kim, Cheol-Soo;Park, Soon-Ki
    • 한국식물생명공학회:학술대회논문집
    • /
    • 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
    • /
    • pp.95-105
    • /
    • 2005
  • Transposon-mediated insertional mutagenesis provides one of the most powerful tools for functional studies of genes in higher plants. This project has been performed to develop a large population of insertional mutations, and to construct databases of molecular information on Ds insertion sites in rice. Ultimate goals are to supply genetic materials and information to analyze gene function and to identify and utilize agronomically important genes for breeding purpose. Two strategies have been employed to generate the large scale of transposon population in a Japonica type rice, Dongjin Byeo; 1) genetic crosses between Ac and Ds lines and 2) plant regeneration from seeds carrying Ac and Ds. Our study showed that over 70% of regenerated plants generally carried independent Ds elements and high activity of transposition was detected only during regeneration period. Ds-flanking DNA amplified from leaf tissues of F2 and T1 (or T2) plants have been amplified via TAIL-PCR and directly sequenced. So far, over 65,000 Ds lines have been generated and over 9,500 Ds loci have been mapped on chromosomes by sequence analysis. Database of molecular information on Ds insertion sites has been constructed, and has been opened to the public and will be updated soon at http://www.niab.go.kr. Detailed functional analysis of more than 30 rice mutants has been performed. Several Ds-tagged rice genes that have been selected for functional analysis will be briefly introduced. We expect that a great deal of information and genetic resources of Ds lines would be obtained during the course of this project, which will be shared with domestic and international rice researchers. In addition to the Japonica rice, we have established the tagging system in an rice line of indica genetic background, MGRI079. MGRI079 (Indica/Japonica) was transformed with Agrobacteria carrying Ac and Ds T-DNA vectors. Among transgenic lines, we successfully identified single-copy Ds and Ac lines in MGR1079. These lines were served as ‘starter lines’ to mutagenize Indica genetic background. To achieve rapid, large scale generation of Ds transposant lines, MGR1079 transformants carrying homozygous Ac were crossed with ones with homozygous Ds, and $F_2$seeds were used for plant regeneration. In this year, over 2,000 regeneration plants were grown in the field. We are able to evaluate the tagging efficiency in the Indica genetic background in the fall.

  • PDF

Transgenic cucumber expressing the 54-kDa gene of Cucumber fruit mottle mosaic virus is highly resistance and protect non-transgenic scions from soil infection

  • Gal-On, A.;Wolf, D.;Antignus, Y.;Patlis, L.;Ryu, K.H.;Min, B.E.;Pearlsman, M.;Lachman, O.;Gaba, V.;Wang, Y.;Yang. J.;Zelcer, A.
    • 한국식물병리학회:학술대회논문집
    • /
    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
    • /
    • pp.148.2-149
    • /
    • 2003
  • Cucumber fruit mottle mosaic tobamovirus (CFMMV) causes severe mosaic symptoms with yellow mottling on leaves and fruits, and occasionally severe wilting of cucumber plants. No genetic source of resistance against this virus has been identified. The genes coding for the coat protein or the putative 54-kDa replicase were cloned into binary vectors under control of the SVBV promoter. Agrobacterium-mediated transformation was peformed on cotyledon explants of a parthenocarpic cucumber cultivar with superior competence for transformation. R1 seedlings were evaluated for resistance to CFMMV infection by lack of symptom expression, back inoculation on an alternative host and ELISA. From a total of 14 replicase-containing R1 lines, 8 exhibited immunity, while only 3 resistant lines were found among a total of 9 CP-containing lines. Line 144 homozygous for the 54-kDa replicase was selected for further resistance analysis. Line 144 was immune to CFMMV infection by mechanical and graft inoculation, or by root infection following planting in CFMMV-contaminated soil. Additionally, line 144 showed delay of symptom appearance following infection by other cucurbit-infecting tobamoviruses. Infection of line 144 plants with various potyviruses and cucumber mosaic cucumovirus did not break the resistance to CFMMV. The mechanism of resistance of line 144 appears to be RNA-mediated, however the means is apparently different from the gene silencing phenomenon. Homozygote line 144 cucumber as rootstock demonstrated for the first time protection of a non-transformed scion from soil inoculation with a soil borne pathogen, CFMMV.

  • PDF

Genotypes of commercial sweet corn F1 hybrids

  • Kang, Minjeong;Wang, Seunghyun;Chung, Jong-Wook;So, Yoon-Sup
    • 한국작물학회:학술대회논문집
    • /
    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
    • /
    • pp.107-107
    • /
    • 2017
  • Sweet corns are enjoyed worldwide as processed products and fresh ears. Types of sweet corn are based on the gene(s) involved. The oldest sweet corn type has a gene called "sugary (su)". Sugary-based sweet corn was typically named "sweet corn". With its relatively short shelf life and the discovery of a complementary gene, "sugary enhanced (se)", the sweet corn (su only) was rapidly replaced with another type of sweet corns, sugary enhanced sweet corn, which has recessive homozygous su/su, se/se genotype. With the incorporation of se/se genotype into existing su/su genotype, sugary enhanced sweet corn has better shelf life and increased sweetness while maintaining its creamy texture due to high level of water soluble polysaccharide, phytoglycogen. Super sweet corn as the name implies has higher level of sweetness and better shelf life than sugary enhanced sweet corn due to "shrunken2 (sh2)" gene although there's no creamy texture of su-based sweet corns. Distinction between sh2/sh2 and su/su genotypes in seeds is phenotypically possible. The Involvement of se/se genotype under su/su genotype, however, is visually impossible. The genotype sh2/sh2 is also phenotypically epistatic to su/su genotype when both genotypes are present in an individual, meaning the seed shape for double recessive sh2/sh2 su/su genotype is much the same as sh2/sh2 +/+ genotype. Hence, identifying the double and triple recessive homozygous genotypes from su, se and sh2 genes involves a testcross to single recessive genotype, chemical analysis or DNA-based marker development. For these reasons, sweetcorn breeders were hastened to put them together into one cultivar. This, however, appears to be no longer the case. Sweet corn companies began to sell their sweet corn hybrids with different combinations of abovementioned three genes under a few different trademarks or genetic codes, i.g. Sweet $Breed^{TM}$, Sweet $Gene^{TM}$, Synergistic corn, Augmented Supersweet corn. A total of 49 commercial sweet corn F1 hybrids with B73 as a check were genotyped using DNA-based markers. The genotype of field corn inbred B73 was +/+ +/+ +/+ for su, se and sh2 as expected. All twelve sugary enhanced sweet corn hybrids had the genotype of su/su se/se +/+. Of sixteen synergistic hybrids, thirteen cultivars had su/su se/se sh2/+ genotype while the genotype of two hybrids and the remaining one hybrid was su/su se/+ sh2/+, and su/su +/+ sh2/+, respectively. The synergistic hybrids all were recessive homozygous for su gene and heterozygous for sh2 gene. Among the fifteen augmented supersweet hybrids, only one hybrid was triple recessive homozygous (su/su se/se sh2/sh2). All the other hybrids had su/su se/+ sh2/sh2 for one hybrid, su/su +/+ sh2/sh2 for three hybrids, su/+ se/se sh2/sh2 for three hybrids, su/+ se/+ sh2/sh2 for four hybrids, and su/+ +/+ sh2/sh2 for three hybrids, respectively. What was believed to be a classic super sweet corn hybrids also had various genotypic combination. There were only two hybrids that turned out to be single recessive sh2 homozygous (+/+ +/+ sh2/sh2) while all the other five hybrids could be classified as one of augmented supersweet genotypes. Implication of the results for extension service and sweet corn breeding will be discussed.

  • PDF

Analysis of genome variants in dwarf soybean lines obtained in F6 derived from cross of normal parents (cultivated and wild soybean)

  • Roy, Neha Samir;Ban, Yong-Wook;Yoo, Hana;Ramekar, Rahul Vasudeo;Cheong, Eun Ju;Park, Nam-Il;Na, Jong Kuk;Park, Kyong-Cheul;Choi, Ik-Young
    • Genomics & Informatics
    • /
    • 제19권2호
    • /
    • pp.19.1-19.9
    • /
    • 2021
  • Plant height is an important component of plant architecture and significantly affects crop breeding practices and yield. We studied DNA variations derived from F5 recombinant inbred lines (RILs) with 96.8% homozygous genotypes. Here, we report DNA variations between the normal and dwarf members of four lines harvested from a single seed parent in an F6 RIL population derived from a cross between Glycine max var. Peking and Glycine soja IT182936. Whole genome sequencing was carried out, and the DNA variations in the whole genome were compared between the normal and dwarf samples. We found a large number of DNA variations in both the dwarf and semi-dwarf lines, with one single nucleotide polymorphism (SNP) per at least 3.68 kb in the dwarf lines and 1 SNP per 11.13 kb of the whole genome. This value is 2.18 times higher than the expected DNA variation in the F6 population. A total of 186 SNPs and 241 SNPs were discovered in the coding regions of the dwarf lines 1282 and 1303, respectively, and we discovered 33 homogeneous nonsynonymous SNPs that occurred at the same loci in each set of dwarf and normal soybean. Of them, five SNPs were in the same positions between lines 1282 and 1303. Our results provide important information for improving our understanding of the genetics of soybean plant height and crop breeding. These polymorphisms could be useful genetic resources for plant breeders, geneticists, and biologists for future molecular biology and breeding projects.

Expression of yeast Hem1 gene controlled by Arabidopsis HemA1 promoter improves salt tolerance in Arabidopsis plants

  • Zhang, Zhi-Ping;Yao, Quan-Hong;Wang, Liang-Ju
    • BMB Reports
    • /
    • 제43권5호
    • /
    • pp.330-336
    • /
    • 2010
  • 5-Aminolevulinate (ALA) is well-known as an essential biosynthetic precursor of all tetrapyrrole compounds, which has been suggested to improve plant salt tolerance by exogenous application. In this work, the gene encoding aminolevulinate synthase (ALA-S) in yeast (Saccharomyces cerevisiae Hem1) was introduced into the genome of Arabidopsis controlled by the Arabidopsis thaliana HemA1 gene promoter. All transgenic lines were able to transcribe the YHem1 gene, especially under light condition. The chimeric protein (YHem1-EGFP) was found co-localizing with the mitochondria in onion epidermal cells. The transgenic Arabidopsis plants could synthesize more endogenous ALA with higher levels of metabolites including chlorophyll and heme. When the $T_2$ homozygous seeds were cultured under NaCl stress, their germination and seedling growth were much better than the wild type. Therefore, introduction of ALA-S gene led to higher level of ALA metabolism with more salt tolerance in higher plants.