• Title/Summary/Keyword: tissue-culture

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Propagation of Bell Pepper (Capsicum annuum var. grossum) through tissue culture

  • Senarath, WTPSK;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.131-132
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    • 2003
  • Leaf discs and apical meristems were cultured in Murashige and Skoog (MS) medium supplemented with cytokinin and auxin at different concentrations. Callus production was observed in all tested media after six days of incubation. Callus produced in the presence of high concentration of NAA (2.0mg/1) was fragile in texture and yellow in colour. Highest callus formation was observed from leaf discs in the medium supplemented with 1.0mg/1 NAA and 0.5 mg/l BAP in dark at $25{\pm}1{\circ}C$. Percentage of callus formation was 95% and mean callus fresh weight was 654.88 43.53 mg. Shoots were induced from the callus after 4 weeks in 1/2MS medium supplemented with BAP and kinetin both at 0.5mg/1. When elongated shoots were separated and transferred into multiplication medium (MS+0.5mg/1 BAP+0.5mg/1 kinetin) multiplication rate was 6.4 after 6 weeks. Higher concentrations of BAP caused callus production at the base. Direct shoot induction was observed from apical meristems in MS medium in the presence of 0.175 mg/1 IAA + 2.25mg/1 BAP and 0.175 mg/1 IAA + 3.0 mg/1 BAP in 16 hour day at $25{\pm}1{\circ}C$. Explants (apical meristems) elongated to form a single shoot forming a callus at the base. Adventitious buds were sprouted out from the base. Percentage explants which producing shoots was 28.57 and 65.5 respectively. Multiple shoot induction was also observed in the same media. Highest multiple shoot production was observed in the presence of 0.175 mg/l IAA and 3.0mg/l BAP, Mean number of shoots per explant was 5.36 and the mean shoot length was $16.66{\pm}4.15$mm. Shoots (20 30m length) were tested for root induction. Excised shoots were transferred into rooting media, which contains different concentrations of NAA and IAA. Best rooting performance was observed in 1/2MS medium supplemented with 0.1mg/1 NAA after 10 days of incubation in 16 hr photoperiod at $25{\pm}1{\circ}C$. Mean number of roots per shoot was 6 and the mean root length was 252mm. Rooted plantlets were transferred into sterile coir dust:sand (1:4) mixture and maintained in a humid chamber for two weeks, They were gradually exposed to the natural environment. After three weeks they were transferred to pots containing coir dust:sand (1:2) mixture for further development where the 90% survival was observed.

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Evaluation of Drought Stress in Native Kimchi Cabbage from Korea (국내 재래종 배추 유전자원의 내건성 평가)

  • Si-Hong Kim;Soo-Yong Choi;Eui-Sub Lee;Young-Hyun Woo;Jae-Yun Heo
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.69-69
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    • 2020
  • 식물체가 건조 스트레스를 받으면 각 기관 물질 생산의 변이, 분화 및 발달 억제를 통해서 식물의 생산활동을 현저히 저하시켜, 식물의 생장, 형태, 개체발생 및 대사생리에 영향을 미치는 것으로 연구되어 왔다. 최근 기후 온난화로 인해 온도가 점진적으로 상승하고 가뭄과 같은 이상기상이 빈번하게 발생함에 따라 많은 노지 작물의 농업 생산성이 약화되고 있다. 배추는 우리나라의 대표적인 식품 중 하나인 김치의 주재료로 연중 안정적인 생산과 공급이 필요시 되지만, 배추의 경우 건조 조건에서 엽육조직의 붕괴와 같은 생리장해가 발생되기 때문에 최근 발생되고 있는 이상 기후의 영향으로 안정적인 생산이 어려워지고 있는 실정이다. 하지만, 배추에서는 이상 기후에 대응이 가능한 내건성 품종의 육성을 위한 연구는 미흡한 실정이다. 따라서 본 연구에서는 내건성이 높은 배추 개발을 위한 기초소재를 찾는 것을 목적으로 농촌진흥청 유전자원센터에서 보존하고 있는 재래종 배추 30계통을 분양받아 강릉원주대학교 생명과학대학 내의 조직배양실을 이용하여 연구를 수행하였다. 배지는 Tissue Culture Square Dish(125×125×20mm)에 Agar를 녹여 40ml씩 분주하여 고체배지를 조성한 후, 건조 처리구 Polyrthylene glycol 6000(PEG) 0%(Control), 20%(Mild Stress), 30%(Severe Stress)를 설정하여 60ml씩 추가 분주하여 배양기 28℃에서 15시간 처리를 하였다. 분양받은 각 계통의 종자는 1% 차아염소산 나트륨으로 10분간 종자표면을 살균한 후, 5번 정도 멸균수로 헹군 후, 표면 살균한 재래종 종자를 고체배지 시험관에 6립씩 치상하였다. 식물체 생육은 각 처리구별 3반복으로 하였으며 주간 12시간 주기, 광도 2,400Lux, 온도 20℃의 조직배양실에서 치상 후 7일간 생육하였다. 치상 후 7일간 생육시킨 식물체를 채취하여 지상부생체중, 지하부 생체중, 뿌리 길이를 측정하였으며, 지상부와 지하부로 나누어 50℃에서 72시간 건조시킨 후, 건물 생산량을 조사하였다. 본 실험 결과 건조 처리(PEG-6000)는 배추의 생장을 저해하였지만, 생육 저해 정도는 계통간의 차이가 있는 것을 관찰할 수 있었으며 무처리구와 스트레스 처리구간의 생장량 변화 정도를 기반으로 군집분석을 수행한 결과 'IT110483'과 'IT104903' 계통이 실험에 공시된 계통들 중 상대적으로 강한 건조 내성을 가진 것을 확인할 수 있었다. 따라서, 재래종 배추 계통 중 일부는 건조내성이 강한 새로운 배추 품종을 육성하는데 있어 유용하게 이용될 수 있을 것으로 사료된다.

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Optimization of in vitro lily culture system with different treatments of taurine (타우린 처리에 의한 나리 기내 식물체 생산체계 최적화)

  • Lee, Sang-Hee;Yang, Hwan-Rae;Kim, Sun Tae;Jun, Tae Hwan;Kim, Yong Chul;Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.484-489
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    • 2017
  • Lilies as cut flowers are one of the most popular ornamental plants in South Korea. It is necessary to develop lily cultivars with high qualities. Therefore, highly efficient propagation systems are needed following release of elite cultivars. In this study, we used taurine treatment to improve the growth conditions including shoot and bulb formation, fresh weight gain, and reduction of rooting and browning. We experimentally evaluated the effect of taurine as a growth stimulator, at concentrations of 0, 2.5, 5, 10, 15 and 20 mg/l. The results showed that 20 mg of taurine enhanced shoot formation by 85% and increased fresh weight 5.5-fold, which was higher than the approximately four-fold increase in the control. In addition, multiple bulb formation rate was increased by 80% and rooting by 82% following exposure to 20 mg/l of taurine. The efficiency of taurine treatment was higher than that of control with 50% multiple bulb formation rate and 60% rooting rate. The browning was 10.6% at 2.5 mg/l of taurine when compared with 0.8% at 20 mg/l. Taurine showed a positive effect on the overall growth of lily plants in terms of increased fresh weight, shoot formation rate, rooting, and formation of multiple bulbs, indicating that taurine can be used as an alternative to amino acids or as an antioxidant such as citrate and vitamin C in plant tissue culture.

Effect of Medium and Growth Regulators on Tuber Propagation by in vitro Culture of Yam (Dioscorea japonica TUNBERG) (산약(山藥)의 기내증식(器內增殖)에 미치는 배지(培地)와 생장조정제(生長調整濟)의 영향(影響))

  • Lee, Hyo-Sung;Ryu, Su-Nho;Lee, Jung-Il;Cho, Chae-Yun
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.1
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    • pp.28-37
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    • 1993
  • The low seed propagation is one of the problem needed a lot of seed tuber for the propagation in yam. Therefore this experiment was carried out to understand the possiblility of seed tuber propagation by tissue culture of yam. In-vitro stem node of yam was cultured by concentration treatments of 1/2, 1/4 and 1/8 with MS medium additted with each concentration levels of IAA, NAA, IBA, kinetin and BA. Acorrding to the Iower concentration than MS medium, length of shoots was promoted, leaf emergence shoots and rooting shoots were increased at 1/8 MS medium during the culturing period of stem node in yam. Fixed IBA and kinetin under the concentration of MS mdeium was inhibited severely by the heigh concentration additted with lAA $1mg\;/\;{\ell}\;and\;NAA\;4mg\;/\;{\ell}$. But fixed IBA $5mg\;/\;{\el}l\;and\;kinetin\;2mg\;/\;{\ell}$ with concentration of 1/8MS medium was remarkably promoted leaf emergence shoots and rooting shoots by $1mg\;/\;{\ell}$ of additted lAA and NAA. Percentage of induced shoots was increased by combination treatments of lAA. $1.5mg\;/\;{\ell}\;and\;kinetin\;2mg\;/\;{\ell}$, also leaf emergence shoots and rooting shoots were promoted by combination treatments of lAA $1.5mg\;/\;{\ell}\;and\;kinetin\;2mg\;/\;{\ell}$.

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Effect of Phosphate Bio fertilizer Produced by Enterobacter intermedium on Rhizosphere Soil Properties and Lettuce Growth (Enterobacter intermedium으로 제조된 인산생물비료가 토양 특성 및 상추의 생육에 미치는 영향)

  • Park, Bum-Ki;Na, Jung-Heang;Hwang-Bo, Hoon;Lee, In-Jung;Kim, Kil-Yong;Kim, Yong-Woong
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.1
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    • pp.15-24
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    • 2005
  • Enterobacter intermedium oxidizes glucose to gluconic acid and sequentially converts gluconic acid to 2-ketogluconic acid (2-KGA) under aerobic condition. Shaking incubation of E. intermedium in a broth medium containing 22.5 g glucose, 8.2 g gluconic acid and 40 g rock phosphate per liter resulted in $1028mg\;L^{-1}$ soluble phosphate. The culture broth was used as phosphate bio-fertilizer (PBF) in this experiment. To evaluate PBF produced by E. intermedium on lettuce growth, five treatments (PBF1/3, PBF2/3, PBF3/3, BP, and MF) were used. In MF and BP treatments, $P_2O_5$ 5.9 kg of mineral fertilizer per 10a was added, while in PBF1/3, PBF2/3, and PBF3/3 treatments, culture broth containing one third, two third, and same amount of soluble $P_2O_5$ 5.9 kg per 10a was applied, respectively. At 20, 35, and 50 days after transplanting of lettuce, plant growth components, biomass, enzyme activities and soil chemical properties were analyzed. Dehydrogenase activity and available phosphate concentration of rhizosphere in phosphate bio fertilizer treatments (PBF1/3, PBF2/3, PBF3/3) were generally higher compared to MF and BP treatments. Soil biomass in PBF3/3 treatment was significantly higher than MF and BP treatments at early growth stage. However, there was no significant difference among all treatments with time. Plant fresh weights in PBF1/3, PBF2/3, and MF treatments were better than those in BP and PBF3/3 treatments. However, in PBF2/3 treatment the highest fresh weight was discovered where alkaline phosphatase activity was generally higher than other treatments at 35 and 50 days. Enhancement of lettuce growth at 35 and 50 days in PBF2/3 treatment was associated with greater phosphate uptake in lettuce tissue. As regarding all results, PBF showed better lettuce growth compared to mineral phosphate fertilizer where PBF2/3 treatment resulted in increase of lettuce fresh weight by 23% and phosphate uptake by 50%.

1-β-D-Arabinofuranosyl-cytosine Induces Chromosomal Breaks in vitro (In vitro에서 1-β-D-arabinofuranosyl-cytosine의 염색체 파열 유도)

  • Jeon, In-sang
    • Clinical and Experimental Pediatrics
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    • v.46 no.12
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    • pp.1186-1193
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    • 2003
  • Purpose : Fragile sites are points on chromosomes which tend to break non-randomly when exposed to specific chemical agents or conditions of tissue culture. The chromosomal break induced by the antineoplastic drug, 1-${\beta}$-D-arabinofuranosyl-cytosine(Ara-c), was investigated to study the laboratory conditions in which the incidence of chromosomal break could be enhanced. Besides, the fragile sites induced by Ara-C were investigated and compared to the already known locations of the specific chromosomal alterations observed in specific neoplasms. Methods : T-lymphocytes from theree normal males and three females were cultured for 48 hours. Cells from each individual were exposed to the Ara-C for an additional 24 hours. After the caffeine was added during the last six hours culture, the metaphase chromosomes were prepared following the conventional method. A site was considered fragile if it was found to break two or more per 100 chromosomal breaks in more than four of six individuals tested. Results : Ara-C induced 252.1 chromosomal breaks per 100 mitotic cells and this result was significantly higher than that of the control, which induced 25.2 breaks(P<0.05). The incidence of the chromosomal break by Ara-C was higher, if cultured in the MEM-FA, which has no folic acid, than in the RPMI 1640 which contains enough folic acid(P<0.05). The most common break site by Ara-C was 3p14.2(FRA3B). There were 20 fragile sites induced by Ara-C. Among these 20 fragile sites, seven coincided with the locations of the mapped oncogenes, JUN, SKI, REL, N-MYC, FHIT, MET, ETS-1, and FOS. Conclusion : S phase specific chemotherapeutic agent, Ara-C, induced the expression of the chromosomal fragile sites effectively using the T-lymphocyte in vitro. Some of the fragile sites by Ara-C highly coincided with the oncogenes and neoplasm specific chromosome breakpoints. In this regard, the fragile sites reported here could provide the unknown neoplasm related chromosomal alternation points.

Growth and Tissue Nutrient Responses of Fraxinus rhynchophylla, Fraxinus mandshurica, Pinus koraiensis, and Abies holophylla Seedlings Fertilized with Nitrogen, Phosphorus, and Potassium at a Nursery Culture (묘포에서 질소, 인, 칼륨 비료주기가 물푸레나무, 들메나무, 잣나무, 전나무의 생장 및 양분에 미치는 영향)

  • Park, Byung-Bae;Byun, Jae-Kyung;Kim, Woo-Sung;Sung, Joo-Han
    • Journal of Korean Society of Forest Science
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    • v.99 no.1
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    • pp.85-95
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    • 2010
  • The purpose of this study was to quantitatively measure both growth performances and nutrient responses of Fraxinus rhynchophylla, Fraxinus mandshurica, Pinus koraiensis, and Abies holophylla seedlings, which are commercially planted in Korea, to nitrogen, phosphorus, and potassium fertilization. We used Dickson's quality index (QI) to compare growth performances and vector diagnosis to interpret nutrient status. Nitrogen fertilization increased more height and root collar diameter growth in F. rhynchophylla and F. mandshurica relative to no fertilization treatment. The QI of F. rhynchophylla and F. mandshurica was the highest on N treatment, but there were no significant differences between treatments for P. koraiensis and A. holophylla. Nitrogen fertilization increased total dry weight by 43, 41, 26, -9% for F. rhynchophylla, F. mandshurica, P. koraiensis and A. holophylla, respectively. In F. rhynchophylla, N fertilization increased N contents with similar N concentrations ("sufficiency"), decreased both P concentrations and P contents ("antagonism"), and decreased K contents with similar K concentrations ("toxic accumulation"). In P. koraiensis, N fertilization decreased N, P, and K concentrations because of more dry weight increases compared to uptaken contents ("dilution"), but N fertilization decreased N, P, and K contents with similar N, P, and K concentrations ("toxic accumulation"). In the light of quality index and vector diagnosis, F. rhynchophylla and F. mandshurica seedlings treated with N fertilization would have high field performance.

Effect of $CO_2$ Enrichment on the Differentiation of Multi-shoots and Saponin contents in Tissue culture of Korean ginseng (Panax ginseng C. A. Meyer) (인삼(人蔘) 조직배양(組織培養)에서 $CO_2$처리(處理)가 multi-shoot 분화(分化) 및 사포닌 함량(含量)에 미치는 영향(影響))

  • Chung, Chan-Moon;Bae, Kil-Kwan
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.4
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    • pp.296-302
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    • 1999
  • This experiment was conducted to study the effect of $CO_2$(0, 2, 500, 5, 000, 10, 000ppm) enrichment by enabling ventilation on micropropagation of multi-shoot and on the saponin contents in vitro in Korean ginseng (Panax ginseng C. A. Meyer). Embryo was cultured in Murashige and Skoog medium added 3mg/ l of Indolbutyric acid, Benzyladenin and Gibberellic acid $(GA_3)$, respectively. $CO_2$, enrichment had little effects on the number of adventitious buds and shoots originated from adventitious buds. The ratio of differentiated shoots to adventitious buds were about 50% in $CO_2$, enrichment treatment. The shoots originated from adventitious bud showed more rapid growth and had larger leaf area than the shoots originated from the leaf primordia did. The number of shoot primordia was the highest in 2, 500ppm of $CO_2$ enrichment treatment. On the contrary, 10,000ppm of $CO_2$, enrichment made smaller the number of shoot primordia and ratio of shoots to shoot primordia. The range of shoots differentiated was from shoot primordia were 15. 4 to 23. 9. The rate of dry weight of cultured shoots showed lowest (7. 5%) in control and highest (8. 59%) in 2, 500ppm of $CO_2$, enrichment. Rate of in vitro flower in control was 7.6% and that in 2500ppm of $CO_2$ was about twice (15.7-16.3%) as much as in control. Flower number per a embryo cultured was about 1.2-1.3. In the multi-shoots with callus enriched by 2, 500ppm of $CO_2$, the contents of crude saponin and ginsenosides in multi-shoots alone were higher than in multi-shoots with callus. The characteristics of ginsenosides in multi-shoots were especially the higher content of ginsenoside Rd, Re, and $Rg_1$.

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Studies on the Contents of the Nutrients and the Nucleotides in Soup Stock During Cooking of Cow's Rumen-Reticulum (소양(Cow's Rumen-reticulum)영양성의 가열조리시 분과 맛성분의 용출에 관한 연구)

  • Lee, Yeon-Sook;Park, Dong-Yeon;Park, Jung-Sook
    • Journal of the Korean Society of Food Culture
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    • v.4 no.3
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    • pp.245-251
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    • 1989
  • To estimate the nutritional values and the optimum cooking method of the Cow's rumen-reticulum (tripe) which is a Korean traditional food, the nutrients of raw tissue of tripes, the changes in total N, ${\alpha}-amino$ N, calcium and phosphorus contents and nucleotides and their related compounds contents in soup stock prepared of tripe tissues according to various boiling time period with saucepot or pressure cooker and various ratios of the water to the tripe (wt/wt) were measured. No significant difference was observed in nutrients contents between rumen (1st stomach) and reticulum (2nd stomach) which contained 83% moisture, 0.4-0.5% ash, 3% fat, 13% protein, 50-56 mg% calcium, 75-76 mg% phosphorus. 75-77 mg% ${\alpha}-amino$ N on fresh weight basis. The results obtained show that the significant loss of nutrients observed after removing epitherial cell layer from tripe, and in aspect of the nutrients contents, the nucletides contents, and the sensory evaluation score in soup stock, the optimum cooking time period of tripe was 8 hours in boiling in saucepot and 1 or 2 hours in cooking in pressure cooker, and the ratio of the water to the tripe (wt/wt) was above ten.

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Elucidation of Mode of Action of Pantoea agglomerans 59-4 for Controlling Garlic Blue Mold (마늘 푸른곰팡이병 방제용 Pantoea agglomerans 59-4의 억제기작 해석)

  • Kim, Yong-Ki;Kwon, Mi-Kyung;Yeh, Wan-Hae;Hong, Sung-Jun;Jee, Hyung-Jin;Park, Jong-Ho;Han, Eun-Jung;Park, Kyung-Seok;Lee, Sang-Yeob;Lee, Seong-Don
    • Research in Plant Disease
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    • v.16 no.2
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    • pp.163-169
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    • 2010
  • To screen for potential biocontrol agents against postharvest disease of garlics caused by Penicillium hirsutum, a total of 1292 isolates were isolated from the rhizoshere or rhizoplane of Allium species. Among them, S59-4 isolate was selected as a potential biocontrol agent by in vivo wounded garlic bulb assay. The isolate was identified as Pantoea agglomerans (Pa59-4) through Biolog system. Pa59-4 did not inhibit the mycelial growth of P. hirsutum in dual-culture with P. hirsutum on tryptic soy agar. In order to elucidate mode of action of Pa59-4 on biological control, nutrient competition between Pa59-4 and P. hirsutum was investigated by the simple method using tissue culture plates with cylinder inserts containing defusing membrane reported by Janisiewicz et al. (2000). The results showed that Pa59-4 effectively suppressed spore germination and mycelial growth of blue mold in the low concentration (0.5%) of garlic juice, but it did not suppress those of blue mold in the high concentration (5%) of garlic juice. This result suggests that the mechanism in biocontrol of garlic blue mold by Pa 59-4 may be involved in nutrient competition with P. hirsutum on garlic bulbs.