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Epigenetic control of LTR retrotransposons in plant germline and somatic cells

  • Lee, Seung Cho;Parent, Jean-Sebastien;Ernst, Evan;Berger, Frederic;Grimanelli, Daniel;Martienssen, Robert A.
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.20-20
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    • 2017
  • Plant genomes include heterochromatic loci that consist of repetitive sequences and transposable elements. LTR retrotransposon is the major class of transposons in advanced plants in terms of proportion in plant genome. The elements contribute not only to genome size but also to genome stability and gene expression. A number of cases have been reported transposon insertions near genic regions affect crop traits such as fruit pigments, stress tolerance, and yields. Functional LTR retrotransposons produce extrachromosomal DNA from genomic RNA by reverse transcription that takes place within virus-like-particles (VLPs). DECREASED DNA METHYLATION 1 (DDM1) plays important roles in maintaining DNA methylation of heterochromatin affecting all sequence contexts, CG, CHG, and CHH. Previous studies showed that ddm1 mutant exhibits massive transcription of retrotransposons in Arabidopsis, but only few of them were able to create new insertions into the genome. RNA-dependent RNA POLYMERASE 6 (RDR6) is known to function in restricting accumulation of transposon RNA by processing the transcripts into 21-22 nt epigenetically activated small interfering RNA (easiRNA). We purified VLPs and sequence cDNA to identify functional LTR retrotransposons in Arabidopsis ddm1 and ddm1rdr6 plants. Over 20 LTR copia and gypsy families were detected in ddm1 and ddm1rdr6 sequencing libraries and most of them were not reported for mobility. In ddm1rdr6, short fragments of ATHILA gypsy elements were detected. It suggests easiRNAs might regulate reverse transcription steps. The highest enriched element among transposon loci was previously characterized EVADE element. It has been reported that active EVADE element is more efficiently silenced through female germline than male germline. By genetic analyses, we found ddm1 and rdr6 mutation affect maternal silencing of active EVADE elements. DDM1-GFP protein accumulated in megaspore mother cell but was not found in mature egg cell. The fusion protein was also found in early embryo and maternal DDM1-GFP allele was more dominantly expressed in the embryo. We observed localization of DDM1-GFP in Arabidopsis and DDM1-YFP in maize and found the proteins accumulated in dividing zone of root tips. Currently we are looking at cell cycle dependency of DDM1 expression using maize system. Among 10 AGO proteins in Arabidopsis, AGO9 is specifically expressed in egg cell and shoot meristematic cells. In addition, mutation of AGO9 and RDR6 caused failure in maternal silencing, implying 21-22 nt easiRNA pathway is important for retrotransposon silencing in female gametophyte or/and early embryo. On the other hand, canonical 24 nt sRNA-directed DNA methylation (RdDM) pathways did not contribute to maternal silencing as confirmed by this study. Heat-activated LTR retrotransposon, ONSEN, was not silenced by DDM1 but the silencing mechanisms require RdDM pathways in somatic cells. We will propose distinct mechanisms of LTR retrotransposons in germline and somatic stages.

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Epigenetic control of LTR retrotransposons in plant germline and somatic cells

  • Lee, Seung Cho;Parent, Jean-Sebastien;Ernst, Evan;Berger, Frederic;Grimanelli, Daniel;Martienssen, Robert A.
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.97-97
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    • 2017
  • Plant genomes include heterochromatic loci that consist of repetitive sequences and transposable elements. LTR retrotransposon is the major class of transposons in advanced plants in terms of proportion in plant genome. The elements contribute not only to genome size but also to genome stability and gene expression. A number of cases have been reported transposon insertions near genic regions affect crop traits such as fruit pigments, stress tolerance, and yields. Functional LTR retrotransposons produce extrachromosomal DNA from genomic RNA by reverse transcription that takes place within virus-like-particles (VLPs). DECREASED DNA METHYLATION 1 (DDM1) plays important roles in maintaining DNA methylation of heterochromatin affecting all sequence contexts, CG, CHG, and CHH. Previous studies showed that ddm1 mutant exhibits massive transcription of retrotransposons in Arabidopsis, but only few of them were able to create new insertions into the genome. RNA-dependent RNA POLYMERASE 6 (RDR6) is known to function in restricting accumulation of transposon RNA by processing the transcripts into 21-22 nt epigenetically activated small interfering RNA (easiRNA). We purified VLPs and sequence cDNA to identify functional LTR retrotransposons in Arabidopsis ddm1 and ddm1rdr6 plants. Over 20 LTR copia and gypsy families were detected in ddm1 and ddm1rdr6 sequencing libraries and most of them were not reported for mobility. In ddm1rdr6, short fragments of ATHILA gypsy elements were detected. It suggests easiRNAs might regulate reverse transcription steps. The highest enriched element among transposon loci was previously characterized EVADE element. It has been reported that active EVADE element is more efficiently silenced through female germline than male germline. By genetic analyses, we found ddm1 and rdr6 mutation affect maternal silencing of active EVADE elements. DDM1-GFP protein accumulated in megaspore mother cell but was not found in mature egg cell. The fusion protein was also found in early embryo and maternal DDM1-GFP allele was more dominantly expressed in the embryo. We observed localization of DDM1-GFP in Arabidopsis and DDM1-YFP in maize and found the proteins accumulated in dividing zone of root tips. Currently we are looking at cell cycle dependency of DDM1 expression using maize system. Among 10 AGO proteins in Arabidopsis, AGO9 is specifically expressed in egg cell and shoot meristematic cells. In addition, mutation of AGO9 and RDR6 caused failure in maternal silencing, implying 21-22 nt easiRNA pathway is important for retrotransposon silencing in female gametophyte or/and early embryo. On the other hand, canonical 24 nt sRNA-directed DNA methylation (RdDM) pathways did not contribute to maternal silencing as confirmed by this study. Heat-activated LTR retrotransposon, ONSEN, was not silenced by DDM1 but the silencing mechanisms require RdDM pathways in somatic cells. We will propose distinct mechanisms of LTR retrotransposons in germline and somatic stages.

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Plant Regeneration via in Vitro Culture of Ovule Obtain by Intergeneric Crossing Between Citrus junos Sieb. et Tanaka and Poncirus trifoliata Raf. (유자와 탱자의 속간교잡후 배주배양에 의한 식물체 유기)

  • 이만상;남궁승박
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.317-322
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    • 1995
  • As a basic research for breeding new varieties, reciprocal -intergeneric crosses between Citrus junos and P.trifoliata were made. F$_1$ hybrid production using in vitro ovule culture, gametogenesis, and fertilization phenomena were investigated. Frequency of fruit set resulting from crossing of Citrus junos and Poncirus Trifoliata was 16.6% while that of Poncirus Trifoliata and Citrus junos was 11.7%. Callus formation occurred well when ovules at the 6th week after pollination were cultured on MT (Murashige and Tucker) medium supplemented with zeatin 0.5 mg/L and NAA 1.0 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4-D 0.1 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4 D 0.1 or 0.5 mg/L. The invitro germination rates of 20-week-old ovules set C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 54.5% and 48.6%, respectively. The emergence ratios of trifoliate hybrids obtained by C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 56.7% and 100%, respectively. The chromosome number of C. junos and P. Trifoliata was n = 9 or 2n = 18, and the sizes of their pollen grain were 33.75 $\mu$ and 25.0 $\mu$. The length and width of embryo sac in C. junos and P. Trifoliata were 69.38~79.23 $\mu$ and 27.50~38.56 $\mu$, and those of egg cells were 17.50~41.50 $\mu$ and 6.25~8.12$\mu$. Fertilization of C. junos and P. trifoliata terminated 72 h after pollination.

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농업용수의 수온 상승에 관한 연구

  • Hwang, Eun;Kim, Cheol-Gyu;Lee, Sang-Beom
    • Water for future
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    • v.5 no.2
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    • pp.17-29
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    • 1972
  • The persent study aims at finding out a means of prevention cool spell damages on the hilly areas. The irrigation plots of 24 hour stored water warm water way and warm water plots, cool water way are respectively established to find out water temperature and influnce on the growing rice plants. The results obtained are summed up as follows. 1. Warm water areas consisted of $5 m^2 Q=0.93 1{\ell}/sec$, V=31 cm/sec, S=1/1, 000, L=81.6m, B=5cm, h=6cm, t=4min 33sec, drops=9 areas, are constructed to help the water temperature of $14.5^{\circ}C$ rise to that of $21.6^{\circ}C$. This indicates lower temperature than $23^{\circ}C$ of critical water temperature in irrigation facilities by $1.45^{\circ}C$ and than $26.2^{\circ}C$ of balanced water temperature of Seoul arears by $4.6^{\circ}C$. But this does not give much influance on rice plant cultivation. 2. The rising of water temperature is influened according to the temperature, solar radiation but the water temperature changes according to the heat absorption of organized materials, weather and terraces. The difference of water temperature could be found in the first growing stage. 3. Through the warm water way of water rises to the temperature of $21.6^{\circ}C$ which also rises to the temperature of around $30^{\circ}C$ in the paddy field of submerged irrigation. The rice plants are comparatively free from prolonged cool damage, reproduction abstructive damage. 4. The water temperature in rice field in proportion to temperature influence of weather condition but the water temperature approaches to that of weather in the days of later growing stage and water temperature become lower than the air temperature in the fruit stage. 5. The water in the submreged field is $10^{\circ}C$ warmer than in the warm water way during the first growing stage period but the water temperature in the warm water way is warmer in the later growing stage period. The cool water of $14.5^{\circ}C$ is warmed to $30.1^{\circ}C$ and rice plants cultivation is free from other damages. 6. The 12% increased production or 570.98kg/10a is made cool water plot by rising the temperature of water from $14.5^{\circ}C$ to $21.6^{\circ}C$ making the water run through warm water way. 7. The damage inflicted by the cool water irrigation during the first growing stage period is the obstruction of peak tillering stage and the obstruction of heading the later growing stage period and the obstruction of fruiting and number of panides per fill.

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Change in the Seed Characteristics and Germination Properties of Ulmus davidiana var. Japonica According to Seed Collection Time (채취 시기에 따른 느릅나무의 종자 형질 및 발아 특성 변화)

  • Tak, Woo Sik;Choi, Chung Ho;Kim, Tae Su
    • Journal of Korean Society of Forest Science
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    • v.95 no.3
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    • pp.316-322
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    • 2006
  • This study was conducted to examine the changes of seed characteristics and germination properties and to determine the most suitable collection time in Ulmus davidiana var. japonica. Seeds were collected six times with the interval of seven days after April 21, 2005 at Jeong-seon. There were differences in seed characteristics and germination properties among seed collection times. The sizes of seed and fruit increased with the collection time, but seed moisture contents decreased with the collection time after increasing until May 5. The 1,000 seeds weight decreased with the collection time after increasing until May 12. Collection time showed high positive correlations (r=0.790, p<0.01) with seed size and negative correlations (r=-0.919, p<0.01) with moisture contents. Percent germination (PG) presented the highest value in seeds collected at May 5, and decreased with the collection time. Mean germination time(MGT), germination speed(GS), and germination performance index(GPI) also showed the highest values in seeds collected at May 5. PG, MGT, and GS had correlations with other seed characteristics except the 1,000 seed weight. The relative growth rate of height and root collar diameter of seedlings presented the highest values in seedling germinated from seed collected at May 5, and seedlings germinated from seeds collected at May 5 and 12 showed the highest values in chlorophyll contents. Consequently it will be possible to product a number of seedlings when the seeds of U. davidiana var. japonica are collected just before drying morphologically.

Effect of Elevated CO2 Concentration and Temperature on the Growth and Ecophysiological Responses of Ginseng (Panax ginseng C. A. Meyer) (CO2농도와 온도증가에 따른 인삼의 생육 및 생리.생태학적 반응 연구)

  • Lee, Kyoung-Mi;Kim, Hae-Ran;Lim, Hoon;You, Young-Han
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.2
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    • pp.106-112
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    • 2012
  • In order to understand the growth and ecophy -siological response of ginseng to global warming condition, we cultivated one and two year ginseng seedlings in control (ambient $CO_2$ + ambient temperature) and global warming treatment (elevated $CO_2$ + elevated temperature) from March 2010 to July 2011. Shoot appearance and initiation of flowering were advanced by 3-4 days in global warming treatment than in control. However, timing of fruit setting and seed ripeness was similar in both control and global warming treatment. Shoot length was longer in global warming treatment than in control, and also the number of leaves was much in global warming treatment. Fresh root weight was not different between control and global warming treatment. Photosynthetic rate was higher in global warming treatment than at control. Photosynthetic rate and transpiration rate were higher in two year seedlings than in one year seedlings at control, but was not different between seedling age of ginseng in global warming treatment. Water use efficiency was higher in one year seedlings than two year seedlings at control and global warming treatment. These results demonstrated that Korean ginseng more or less positively responds to global warming situation.

Screening for Resistance to Downy Mildew among Major Commercial Cucumber Varieties (주요 오이 품종의 노균병에 대한 저항성 검정)

  • Lee, Jung-Sup;Han, Kyung-Sook;Lee, Seong-Chan;Soh, Jae-Woo
    • Research in Plant Disease
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    • v.19 no.3
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    • pp.188-195
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    • 2013
  • This study was carried out for the downy mildew resistant test between 2010 and 2012. A set of 22 accessions belonging to 2 wild species and 20 varieties of the genus Cucumis, originating mainly from the Asian Vegetable Research and Development Center (AVRDC) Gene Centre, was evaluated for resistance to Pseudoperonospora cubensis, causal agent of cucumber downy mildew. The youngest fully expanded true leaves were found suitable for in vitro screening. Both leaf discs and full leaves could be kept fresh longer when applying 0.2 ${\mu}g/ml$ of gibberellin acid (GA). The incubation temperature of $20^{\circ}C$ was found to be the most suitable temperature for symptom development comparing with 15 and $25^{\circ}C$. Symptom development was faster when contact diseased leaf discs (2 weeks after inoculation) on to fresh leaf samples comparing with using conidia suspension ($10^5$ spores/ml). The numbers of spots in 'C-19' were lower than other varieties. 'C-19' variety was also showed the highest level of downy mildew resistant at $20^{\circ}C$ chamber in 6 days after inoculating with pathogen and displayed 0.90 (under 10%) of the infected rate. However, other varieties displayed susceptible in the pathogen sprayed plots. 'C-19' was the most resistant variety and no lesion was observed. Based on all data, 'C-19' can be a useful variety for the prevention of downy mildew.

Correlation between the Dieback Ratio and Cultivation Environment for Apple Orchards Infected by Soil-Borne Diseases in Chungbuk Province (충북에서 사과 주요 토양병에 의한 고사율과 재배환경과의 상관관계)

  • Lee, Sung-Hee;Kwon, Yeuseok;Shin, Hyunman;Kim, Ik-Jei;Nam, Sang-Young;Hong, Eui Yon;Kim, Daeil;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.23 no.1
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    • pp.49-55
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    • 2017
  • The previous study showed that die-back of apple trees caused by soil-borne diseases was significantly high in the apple orchards in Chungbuk province. The correlation between dieback ratio and cultivation environment in apple orchards infected by soil-borne diseases was investigated in this study. The dieback ratio of five orchards diseased by violet root rot and five places infected by white root rot showed significantly positive correlation with Ca content and available $P_2O_5$ content in soil, respectively. Whereas, the dieback ratio of fourteen orchards diseased by Phytophthora root rot was not significant. Subgrouping of cultivation environment analysis showed that the slope degree of orchard and the number of fruit setting also affected the dieback ratio caused by violet root rot and Ca content in soil also affected the dieback ratio caused by white root rot. It showed that the slope degree, soil texture, Mg and Ca content affected the dieback ratio caused by Phytophthora root rot. These results can be applied to reduce die-back ratio by the modification cultivation environment for each soil-borne disease.

Effect of Temperature on Pathogen Growth and Damage Analysis of Leaf Spot Disease on Grapevine Caused by Paeudocercospora vitis in Korea (포도나무 갈색무늬병균(Pseudocercospora vitis (Lếv.) Speg.)의 생육에 미치는 온도의 영향과 갈색무늬병 발생 포도나무의 피해해석)

  • Park Jong-Han;Han Kyeong-Suk;Lee Jung-Sup;Seo Sang-Tae;Jang Han-Ik;Kim Heung-Tae
    • Research in Plant Disease
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    • v.12 no.1
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    • pp.10-14
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    • 2006
  • This study was carried out to know damages of the grapevine trees by the disease to the leaf spot from 2000 to 2003. The isolates collected from different varieties and locations were identified as Pseudocercospora vitis ($(L\acute{e}v)$.) Speg. based on the morphological and cultural characteristics. According to the in vitro test, the range of temperatures for the mycerial growth and the conidical germination of the fungus were from $10^{\circ}C\;to\;35^{\circ}C$ and optimum temperature was $25^{\circ}C$. There were remarkably different features between a low infection trees group and high infection trees group in terms of number of leaves per fruit branch, length and diameter of internode and leaf area.

Effect of Mixed Liquid Fertilization on Growth Responses of Red peppers and Soil Chemical Properties

  • Park, Ji-Suk;Lee, Min-Jin;Lee, Seo-Youn;Kim, Jong-Sung;Lee, Tae-Kyu;Ro, Hee-Myong
    • Korean Journal of Soil Science and Fertilizer
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    • v.48 no.3
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    • pp.225-232
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    • 2015
  • We evaluated the effect of mixed liquid fertilizer (MLF) on growth responses of plants and soil chemical properties. A pot experiment with red pepper (Capsicum annuum L.) using loam soil was conducted for 81 days in a temperature-controlled glasshouse, and four N fertilization treatments were laid out in a completely randomized design with three replicates: control (C), chemical fertilizer treatment (CF), and two rates (MLF-0.5 and MLF-1.0) of MLF treatment. Soils were periodically sampled and analyzed for pH, EC(Electrical Conductivity), total N, inorganic N and total C, and some growth characteristics of red peppers were measured. During the experimental periods, the pH of MLF soils was higher than that of CF soils. Soil EC increased right after application of CF or MLF, and the intial increase persisted in CF and MLF soils at the end of experiment. Soil total-N increased right after application of CF or MLF, and this initial increase persisted only in MLF-1.0 soils. Soil inorganic N content initially increased in CF or MLF-1.0 soils, but the initial increase disappeared in 56 days after transplanting. Soil total-C was maintained higher in MLF-1.0 soils and lower in CF soils than in control soils, and the intial increase in MLF-1.0 soils finally disappeared to the level of control soils. Plant height, dry weight of plant organs (shoots, roots and fruit), and the number, diameter and length of red pepper fruits were greatest in CF plants. On the other hand, the effect of MLF-application was different depending on the rate of application. However, no consistent effect of N treatments on some major elements of the organs of red peppers was observed. The amounts of N taken up by plants were 1.3 g for CF, 0.8 g for MLF-1.0, 0.5 g for MLF-0.5 and 0.4 g for control treatments. The results of this study showed that mixed liquid fertilizer (MLF) could appropriately serve as an alternative to chemical N fertilizer in red pepper cultivation.