보고서 정보
주관연구기관 |
전북대학교 Chonbuk National University |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2015-02 |
과제시작연도 |
2014 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201500010594 |
과제고유번호 |
1395035049 |
사업명 |
국책기술개발 |
DB 구축일자 |
2015-07-11
|
DOI |
https://doi.org/10.23000/TRKO201500010594 |
초록
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Ⅳ. 연구개발결과
<1세부과제: 벼 보급종 안전생산을 위한 채종포 종자전염성 병해충 진단기술 개발>
○ 채종포의 못자리 및 본답에서 키다리병 발병 조사 결과 대부분의 채종포에서 매우 높은 발병률을 보였음
○ 농가 수집 종자를 파종하여 유묘 발병률과 PCR을 이용한 검출률을 비교한 결과 대부분의 수집 종자에서 발병률보다 매우 높은 비율로 키다리병 병원균이 검출되었음 (영농활용: 벼 채종포 종자 키다리병 감염 검사 시 PCR 검정법 이용)
○ 벼 흰잎마름병과 세균벼알마름병 병원균을 시기별로 PCR primer를 이
Ⅳ. 연구개발결과
<1세부과제: 벼 보급종 안전생산을 위한 채종포 종자전염성 병해충 진단기술 개발>
○ 채종포의 못자리 및 본답에서 키다리병 발병 조사 결과 대부분의 채종포에서 매우 높은 발병률을 보였음
○ 농가 수집 종자를 파종하여 유묘 발병률과 PCR을 이용한 검출률을 비교한 결과 대부분의 수집 종자에서 발병률보다 매우 높은 비율로 키다리병 병원균이 검출되었음 (영농활용: 벼 채종포 종자 키다리병 감염 검사 시 PCR 검정법 이용)
○ 벼 흰잎마름병과 세균벼알마름병 병원균을 시기별로 PCR primer를 이용하여 검출한 결과 9월 하순의 황숙기 이후에 조사하였을 때 높은 비율로 검출되어 이 시기가 병원균 검출에 적당한 시기로 판단됨
○ 키다리병 병원균 비산 거리를 조사결과 150m 떨어진 곳에서 채집한 종자에서도 병원균검출(정책건의: 채종포 인접 일반농가에 키다리병 방제를 위하여 보급종자 공급)
○ 프로라츠 유제를 이용한 종자소독 시 PCR primer를 이용한 병원균 검출률이 7일묘에서 27.5%, 15일묘에서 72.5%로 발병률에 비해 매우 높게 나타나 발병이 되지 않았어도 잠복 감염이 되어 있는 것으로 나타났음
<2세부과제: 벼 등숙기 기상 환경 및 가공처리 요인이 종자활력에 미치는 영향 분석 및 종자활력기준 설정>
○ 등숙기간 중 벼 종자가 발아력을 갖는 시기는 출수 후 20~30일, 50% 이상의 종자가 발아될수 있는 시기는 출수 후 35~45일로 품종간 변이를 보임
○ 벼 등숙기간 동안 인공강우 처리 결과, 출수 후 40일 온실처리에서는 호품벼가 3일처리에서 수발아가 발생하였고, 출수 후 50일 온실처리에서는 호품벼가 3일처리부터, 신동진벼는 7일처리부터 수발아가 발생하였음
○ 표준발아검사(25℃) 결과, 85%이상의 발아율을 보인 종자는 입모율(17℃)검사에서 종자 퇴화정도에 따라 큰 차이를 보였으며, 불량환경에서 안정적인 입모확보를 위하여 90% 이상의 표준발아기준 설정이 요구되어진다고 판단됨(영농활용)
○ 수분함량 17% 벼 종자에서 열풍온도처리 70℃의 열풍에서, 수분함량 25%인 종자는 열풍온도처리 60℃ 이상에서 급격히 활력이 떨어지기 시작함. 호품벼가 신동진벼보다 건조온도에 따른 활력감소가 크게 나타남
○ 키다리병의 열풍건조에 의한 방제는 종자수분함량 10% 이내에서 열풍온도 60 ℃ 정도에서 30분 이내로 처리하는 것이 적합할 것으로 판단됨
<3세부과제: 벼 병해 발생 경감을 위한 종자처리 기술개발>
○ 키다리병에 유효한 약제 4점과 미생물 3종을 선정하였음
○ PVP-630, CMC, Xanthan gum 세 종류의 Binder를 이용하여 펠렛팅을 진행한 결과, PVP와 Xanthan gum을 Binder로 사용하는 것이 좋으나, Xanthan gum의 경우 높은 점성을 가지고 있어 Binder로서 부적합하였음
○ 펠렛 종자의 발아율 실험 결과, 초기 일반 종자와 발아율의 차이가 보였으나 11일 경과 일반 종자와 펠렛 종자의 발아율의 차이는 보이지 않았음
○ 종자처리 벼의 야외조건에서의 키다리병 방제효과를 평가한 결과 무처리 대비 Bacillus sp. 코팅종자 처리구에서 낮은 이병률을 보였으며, mancozeb 코팅종자 처리구에서 높은 이병률을 보였음
○ 종자처리 벼의 보관 안정성을 평가한 결과 Bacillus sp. 코팅종자는 온도에 상관없이 모든처리에서 clear zone이 확인되었으며 Fluazinam과 Mancozeb은 코팅하는 과정에서 성질을 잃었을 것으로 판단됨
Abstract
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(Project 1)
In this study, we were investigated to the occurrence of major diseases at the seed production field which was selected the Jeonbuk province.
Non-pathogenic fungi Alternaria alternata, Cladosporium cladosporioides showed the highest detection rate among the seed samples collected i
(Project 1)
In this study, we were investigated to the occurrence of major diseases at the seed production field which was selected the Jeonbuk province.
Non-pathogenic fungi Alternaria alternata, Cladosporium cladosporioides showed the highest detection rate among the seed samples collected in 2012, but pathogenic fungi Bipolaris oryzae, Fusarium fujikuroi , F. graminearum were dectected in 2~8 percents. The results of occurrence of the Bakanae disease of the seedbed was 5 percent at the seed production field and was 15 percent or higher at the farm only the part of Hwangdeung and Sanseo among Jeonbuk area. t the comparison between the detection rate of the pathogen by PCR primer and the incidence of bakanae disease, the detection rate was higher then the incidence. The result of occurrence investigation of bakanae disease of paddy fields of the seed production field were very low incidence at mostly of investigated region but it were 0.5 percent or higher at the part of Hwangdeung, Seosu, and Anseong. In growing period at the seed production fields, Bacterial leaf blight, Bacterial grain rot, and Rice white-tip nematode was not occurred at mostly region of seed production fields in 2013, but the Bacterial leaf blight was very low incidence occurred at the part of Hwangdeung, Yongan, and Deokcheon in 2014, and Bacterial grain rot was very low incidence occurred at the part of Jusan, Taein, Yeongwon, Ongdong in 2013 and 2014.
(Project 2)
The occurrence of frequent extreme weather events due to the recent climate changes have resulted in changes in precipitation and temperature which have a direct impact on crop production. In the case of the rice seed, low temperature in spring and frequent rainfall in autumn could accelerate its decay or delay its germination, resulting in failure in seedling establishment. Therefore, at first, the influence of rainfall during the ripening stage on pre-harvest sprouting (PHS), seed viability, and seed quality was investigated. This research was implemented to investigate pre-harvest sprouting characteristics in two Korean rice cultivars, Shindongjin (SDJ) and Hopum (HP). The panicles of both varieties were sampled after 15 days after heading(DAH) to 60 DAH at intervals of 5 days. As soon as sampled, the panicles were imbibed in water for 14 days at 25℃ to determine the vigor and germination ability of seeds according to location on panicle. To investigate the cause of non-germination of seeds in panicles, tetrazolium test and dormancy breaking were performed. The preharvest sprouting of HP started at 20 DAH while that of SDJ began at 30 DAH. The germination of seeds located in high rank branches within a panicle and the terminal seeds within a branch were earlier and faster and the germination patterns were same in both cultivars. The times at more than 50% of germination in a panicle were 35 DAH (57.0%) in HP and 45 DAH (56.8%) in SDJ. Preharvest sprouting was more than 80% at 50 DAH (82.6% of HP, 92.3% of SDJ) and more than 99% at 60 DAH (99.5%, 99.4%, HP and SDJ). These results indicated that the rate of PHS in a panicle increased with accumulation of the days after heading. The cause of non-germinated seeds at 15-25 DAH in panicle was immature embryo.
After 30 DAH, however, when the non-germinated seeds were hulled, they started to germinate due to dormancy breaking, in which the germination percentage was higher in SDJ than HP. When the rainfall was artificially treated in a greenhouse, HP started to pre-harvest sprouting at three days of rainfall treatment (DRT), but SDJ did not show pre-harvest sprouting at 40 DAH treatment and just 0.3~0.8% at 50 DAH, which was much lower than 15.3~25.8% of HP in the same treatment. After harvest, the seed germination of Hopum decreased about 10~25% compared to non-treated seeds, but that of SDJ decreased much little rate than that of HP. The seed longevity tested by accelerated aging decreased with prolonged rainfall period in both cultivars, but the varietal difference was clear; Shindongjin could withstand longer accelerated aging than HP. Rainfall during the ripening stage induced not only pre-harvest sprouting, but also reduced seed quality and longevity duringstorage, which varied between two cultivars. The seconds, SDJ and HP were treated with an accelerated aging (AA) at 40°C±1 (100 % RH) for 1, 3, 5, 7, 9, 11, 13 and 15 days (d). Each AA seeds was sowed in the soil at 17°C and seedling establishment (SE) of the seeds was evaluated. Various seed tests were carried out to predict SE such as standard germination test (SGT), cool germination test (CGT), cold germination test (CT), seedling growth rate test (SRT). The following activities of antioxidant enzymes tests were also performed: catalase (CAT), peroxidase (POX), superoxide dismutase (SOD), ascorbate peroxidase (APX). Percentage of SE decreased slowly from 75% to 65% by 0 to 3d AA, but it decreased rapidly less than 50% by 5d AA (HP 32 %, SDJ 48 %) and decreased continuously to 0% until 15d AA. The different tests of AA seeds showed various different results. Among the seed tests, SGT (0.889), CGT (0.867), SRT (0.865) and POX (0.856) had high correlation coefficients with SE. In terms of securing SE about 70% under low temperature condition, SGT and POX could be used to predict SE because the two tests were very similar with the change of SE by 0-5 d AA. In the experiment, when the germination percentage in SGT and the activity of POX were above 90% and 2.0 U/mg-1, respectively, the SE was more than about 70% in both SDJ and HP. This results suggested that SGT and POX could be used to predict SE in unfavorite environments in rice transplanting cultivation.
The third, after artificially adjusting seed moisture content (SMC) (HP, SDJ) as SMC 17% and 25%, it was investigated to change drying time and seed vigor by temperatures of 40-100℃. To confirm Bakanae Disease (Fusarium fujikuroi) reduction in rice seeds by hot-air drying treatment during seed drying processing, rice seeds infected with Fusarium fujikuroi were treated at temperatures of 40-100℃ for 5, 10, 15, 20, 25 and 30 minutes. After its treatment, they were placed equidistantly on PDA medium in Petri dishes and incubated at 25°C in the darkness for four days and infected rice seeds were followed by polymerase chain reaction (PCR).Seeds of SMC 17% were dried at 40 to 50℃ for nearly 30 minutes and at 55 to 70℃ for nearly 20 to 15 minutes, and those with Seeds of SMC 25% were dried at 40 to 50℃ for nearly 2 hours, at 50 to 60℃ for nearly 1 hour and 30 minutes and at 65 to 70℃ for 45 to 50 minutes. When the seed vigor changes of the seeds depending on the hot-air drying treatment were investigated, the seeds with Seeds of SMC 17% showed a gradual decrease in vigor, as the temperature rose. Compared to the control, SDJ and HP showed a 95% and 77% decrease in vigor respectively at 70℃ during hot-air drying, but the SDJ seeds with Seeds of SMC 25% showed a rapid fall in vigor at 60℃ and over and most seeds lost vigor at 70℃. Also, the HP began to lose vigor at 45℃ and over and most seeds lost vigor at 70℃. Additionally, the seeds were dried with SMC 12 to 14% and then dry-heat treatment was applied to them at a hot-air drying temperature of 40 to 100℃ to examine if the Bakanae disease was relieved. When the germination (%) was verified after hot-air drying, the germination (%)was over 90% at 6 0℃ for 10 minutes during hot-air drying, but it decreased, starting from 70℃. Likewise, the germination (%) was over 85% by up to 60℃ for 30 minutes, but it decreased, starting from 70℃. These results imply that for prevention of the Bakanae disease, it is good to apply hot-air drying to SMC of less than 10% at 60℃ for about 30 minutes. Furthermore, it is considered that using the PDA medium is hard to verify if the seeds are infected and the PCR-based screening method would be appropriate.
(Project 3)
Consideration should be given to the control of seed-born disease in rice plants to reach to the production of rice crops worldwide. Particularly rice bakanae disease, Fusarium fujukuroi , is one of the serious plant disease and give rise to significant loss of crop production annually. This fungal pathogen exists inside of the seeds as a infective state or sometimes it goes into the seeds from the rice field soil. In the lst stage of R&D (fiscal year 1), infected rice seeds were obtained from a field and infection rates were investigated by seedling in laboratory conditions, followed by polymerase chain reaction (PCR) with the pathogen-specific primers (FM-F: 5'-TAC–GCC–ATC–AGT-GCC–GGA-AC-3'and FM-R: 5'-GAG-ATT-CCG-GCA-CTG-GTG-AC-3'; target size, 383 bp). High rates of infection (>90%) was observed from the investigations. To control the fungal pathogen, 40 commercially available fungicides and 10 useful antifungal bacteria were assayed on agar plate conditions. Inhibition of conidial germination and myelial growth was observed on potato dextrose agar where the chemical fungicides or antibacterial Bacillus subtilis (10 isolates) were treated at a given concentration. Of the test materials, fluazinam (CAS No.: 79622-59-6) and mancozeb (CAS No.: 8018-01-7) showed much stronger antifungal activities compared to the rest. The treatment of B. subtilis KACC 10372, 11994 and 12610 inhibited the growth of hyphal extension on the agar medium. In the second stage of R&D (fiscal year 2), optimization of rice seed pelleting was perofmred at a variety of formulations and the control efficacy of a seed pelleting was investigated in a rice-potting assay system. First to determine the morphology or form of the pelleted rice seeds, 5 different pelleting methods was came up with and tried, such as 1) mixing of seeds, binder P and solidifying material A altogether in a bench-scale granulator, 2) Spray of binder P then putting solidifying material A, 3) Dipping of rice seeeds into the binder P solution and integration of solidifying material A, 4) Granual mixing of the binder P and solidifying material A, and 5) pelleting od seeds with small amount of binder P and the material A and followed by the No. 2 method. From the granulation, the No. 2 granulation produced round pelleting form and most of seeds were pelleted in such way without a significant deformation or errors. Next the optimal insertion ratios of solidifying material A and binder P were determined by applying the solidifying material A at 25, 50, 75, 100, 125, 150 and 175 g per 50 ml of dried rice seeds and binder P at 50, 100 and 150% of each solidifying material A. A combination of the powder A (150 g), binder P (150 ml) and rice seed (50 ml) produced the most ideal form (round type) of seed pelleting at the granulator conditions. The optimization experiment was followed by a bench scale granulation using larger amount of rice seeds and an automatic sprayer for the spray of binder P. In the mass production of pelleting seeds, less amount of binder of P was used, possibly because of the self-packing of rice seeds (600 g of powder A, 150 ml of binder P, and 200 ml rice seeds). In addition, CMC and xanthan gum were used as alternative binders to compate the binding ability of binder P at 0.12%, 0.3%, 0.6% and 1.2%. A concentration of 1.2% binder P was determined as the best binder to the solidifying material A. The finally optimized rice seeds showed similarly high germination rates compared to the non-pelleted rice seeds. To test the control efficacy of rice seed pelleting with the selected fungicides and antifungal microorganisms, the following treatments were designed: 1) non-pelleted seeds, 2) pelleting seeds with mancozeb, 3) pelleting seeds with fluazinam, 4) pelleting seeds with B. subtilis KACC10372, 5) pelleting seeds with B. subtilis KACC 11994, 6) seed planting and spray of fluazinam wettable powder, 7) seed planting as a control. All the treatments received the fungal pathogen as an inoculum. Non-inoculated control served as a negative control. The seed pelleting containing fluazinam and mancozeb showed high control efficacy against the rice bakanae disease. In the 3rd stage of R&D (fiscal year 3), the pelleting rice seeds were evaluated in a paddy field and the biological stability was investigated at different temperature of storages. Fluazinam. macozeb and Bacillus subtilis-pelleted rice seeds were planted on rice nursery soil and watered and it was grown for 17 days in a greenhouse before transplanting. One day before the transplanting the fungal pathogen was inoculated at ca. 100 ml (1.25×107 conidia per ml) per square meter of nursery soil. Two and four weeks after the transplanting, the control efficacy was investigated by counting the number of dead plants per spot. The two chemical fungicides-pelleted rice seeds showed any significant control efficacy in the field, but the B. subtilis KACC10372-pelleted rice seeds showed ca. 72% of control efficacy. The pelleting rice seeds were exposured to 20, 30 and 40°C of temperatures for 2, 4, 6 and 8 weeks, which revealed that the two chemical fungicides were unstable at the granulation process (or not evenly homogenized at the pelleting process) and possibly susceptible to the high temperatures but the B. subtilis isolate showed high biological stability, showing antifungal activity against the fungal pathogen. Additionally, an entomopathogenic fungal isolate was integrated into the rice seeds by pelleting and it showed that the pelleting itself had some effect in pest control, but still the potential of the insect pathogenic fungus needs to be more clearly examined. In conlcusion, these results suggest that three possible candidates to control F. fujikuroi were selected from the chemical and microbial stocks and physically stable seed pelleting was optimzed by the series of several granulation experiments, and finally in field conditions B. subtilis KACC10372-pelleted rice seeds showed high potential in the disease control and stability at high temperature conditions of storage. This seed pelleting technology can be used as an advanced research and development system of highly valuable seeds.
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