Pheromone biosynthesis activating neuropeptide (PBAN) produced in the subesophageal ganglion is known to stimulate pheromone production in the pheromone gland. A cDNA isolated from female adult heads of Maruca vitrata encodes 197 amino acids including PBAN, designated as Mvi-PBAN, and four other neuropeptides (NPs): diapause hormone (DH) homologue, α-NP, β-NP and γ-NP. All of the peptides are amidated in their C-termini and shared a conserved motif, FXPR(or K)L-NH2 structure. Mvi-PBAN consists of 35 amino acids as previously reported (Chang and Ramasamy, 2014). RT-PCR analysis revealed that Mvi-PBAN cDNA was expressed in all examined body parts. Nucleotide sequence analysis of RT-PCR products indicated the Mvi-PBAN sequence was identical in all examined body parts of both sexes. These results suggest that Mvi-PBAN expression is maintained in examined stages or tissues.

Pheromone biosynthesis-activating neuropeptide (PBAN), produced in subesophageal ganglion, is known to stimulate pheromone biosynthesis in Plutella xylostella. The pheromone production is more active in the scotophase than in the photophase, which suggests that there may be changes of gene expression in the pheromone glands. To analyze gene expression involving in pheromone biosynthesis for 24 hrs, we performed transcriptomes of pheromone glands which were isolated at every 4 h. Eleven pheromone biosynthesis-related genes, acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), acyl-CoA dehydrogenase, enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, Δ9 desaturase, Δ11 desaturase, fatty acid reductase (FAR), alcohol oxidase, aldehyde oxidase, and aldehyde reductase were identified. Among these genes, ACC, FAS and chain shortening enzymes involving in early stage of pheromone biosynthesis exhibited their highest expression between AM9 and PM5. Desaturases, Δ9 and Δ11, showed the peak of expression at PM1 and AM5 or PM5, respectively. Interestingly, FAR expression was the highest at AM1, active reproductive period. These results suggest that genes involving in pheromone biosynthesis can be sequentially regulated for their biological roles.

The pheromone biosynthesis in Plutella xylostella is stimulated a neuropeptide, pheromone biosynthesis activating neuropeptide which is produced in subesophageal ganglion. The pheromone production is more active in the scotophase than in the photophase, which suggests that there may be changes of gene expression in the pheromone glands. To analyze gene expression related to pheromone biosynthesis, we performed transcriptomes of pheromone glands which were isolated at every 4 h. Eleven pheromone biosynthesis-related genes, acetyl-CoA carboxylase, fatty acid synthase, acyl-CoA dehydrogenase, enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, Δ9 desaturase, Δ11 desaturase, fatty acid reductase, alcohol oxidase, aldehyde oxidase, and aldehyde reductase were identified. Among these genes, the expression of aldehyde reductase and aldehyde oxidase were relatively higher in the scotophase than in photophase, which may affect increase of pheromone production in the scotophase. Expression of signal genes involving in pheromone biosynthesis such as acyl-CoA desaturase, FAR, PBAN receptor, fatty acid transporter and acyl-CoA binding protein did not exhibited any significant difference.

A rapid cold hardening (RCH) and supercooling capacity usually play crucial roles in survival during the overwintering period in the tobacco budworm, Helicoverpa assulta, a freeze-susceptible species. Cryoprotectants such as polyol or sugar affect RCH and maintain fluid status of hemolymph. This study is performed to identify cryoprotectants as a RCH factor in H. assulta. Pre-exposure of H. assulta larvae to 4℃ significantly increased survival at -10℃ in all developmental stages from egg to adult. RCH was dependent on the duration of the pre-exposure period. RCH also significantly enhanced the supercooling capacity. Analysis of cryoprotectants from the hemolymph revealed that the pre-exposure treatment allowed the larvae to accumulate glycerol and trehalose among polyols examined. In addition, unknown materials from 2 peaks were also increased. TIC analysis revealed 3 predicted formulas for unknown materials, C26H24O20S or C3H4N6OS and C22H20O21. Glycerol-3-phosphate dehydrogenase (GPDH) and glycerol 3-phosphatase (G3P) that involving in glycerol biosynthesis were identified from the transcriptome of H. assulta 4th instar larvae. Based on the expression level of transcripts, the expressions of GPDH and G3P were relatively increased when compared to that of the control, suggesting that these genes contribute to overwintering and biosynthesis of glycerol.

The pheromone biosynthesis in Plutella xylostella is more active in the scotophase than in the photophase, which suggests that there may be changes of gene expression in the pheromone glands. To identify genes contributing to change in pheromone production, we analyzed transcriptomes from pheromone glands of both decapitated females in the photophase and normal ones in the scotophase. Comparative analysis were performed with transcriptomes of pheromone glands from non-decapitated (PG) females and decapitated ones for identification and expression of putative genes associated with pheromone biosynthesis pathway. Deep sequencing for mRNAs in the pheromone gland yielded approximately 7.5Gb and totally 17265 transcript were constructed under a homology cutoff of 10-6 Evalue. Genes putatively involved in pheromone biosynthesis were identified such as acetyl-CoA carboxylase, acetyl-CoAdehydrogenase, fatty acid synthase (FAS), desaturases (Δ9 and Δ 11) and fatty acid reductases (FAR) including pgFAR, alcohol oxidase, aldehyde oxidase and aldehyde reductase, etc. Expression of 6 signal genes involving in pheromone biosynthesis such as acyl-CoA desaturase, FAR, PBAN receptor, fatty acid transporter, acyl-CoA binding protein did not exhibited ant significant different in both transcriptomes. Quantitative RT-PCR revealed that expressions of FAS, Δ11 desaturase and pgFAR were higher in PG than that in ΔPG. Based on results, Δ11 desaturase and pgFAR may have a crucial role in sex pheromone biosynthesis of P. xylostella.

현재 재배되고 있는 장미 재배종은 오랜 기간 동안 종간교잡과 종내교잡을 통해 선발되어 온 것으로서 대 부분이 이형잡종의 게놈으로 구성되어 있어 발아율이 저조한 편이다. 따라서, 효율적인 교배를 위해서는 화 분의 활력 검정이 선행되어야 한다. 이에 본 연구에서는 염색액과 발아배지를 이용하여 7품종의 화분 임성율과 발아율 검정하고 이들 결과를 토대로 화분의 크기, 배수 성 및 교배조합과의 상관관계를 알아보고자 실시하였다. 화분 임성율은 ‘스칼렛미미’가 82%로 가장 높았으며 ‘피노키오’가 39%로 가장 낮았다. 발아율 관찰 결과 ‘스칼렛미미’가 41%로 가장 높았으며 ‘미니로사’가 1%로 가장 낮았다. 화분의 크기를 관찰한 결과 화분은 큰 화분 과 작은 화분으로 구분되었으며 화분의 직경은 각각 41.3- 45.4 μm과 30.7-37.4 μm로 관찰되었으며 화분의 크기는 10-40% 정도 차이가 있는 것으로 나타났다. 화분의 크기 와 염색체 전체 길이의 합을 비교한 결과 염색체 크기와 화분의 크기는 정의 상관관계가 있음을 알 수 있었다. 교배조합 검정 결과 착과율과 발아율은 화분 발아율, 임성률 및 배수성과 상관관계가 있음을 확인하였다.

키가 큰 절화용 백합인 ‘Rialto’와 ‘Elodie’ 및 ‘Raizan’ 을 분화 및 화단용으로 사용하기 위하여 ancymidol과 uniconazole 및 paclobutrazol을 농도별로 관주처리 하 였다. 오리엔탈 백합인 ‘Rialto’의 경우, paclobutrazol을 처리한 구에서는 초장뿐만 아니라 잎과 꽃에 있어서도 변화를 관찰할 수 없었다. 그러나 ancymidol 처리구에서 생장은 농도에 관계없이 30% 이상 억제되었다. 한편 엽수나 개화수 등은 무처리구와 비슷한 경향이었다. Uniconazole 1mg·L−1 처리구에서 초장은 무처리구와 비슷한 경향이었으나 5와 10mg·L−1 처리구에서는 30% 정도 단축되었다. 아시아틱 백합인 ‘Elodie’의 경우 ‘Rialto’와 마찬가지로 paclobutrazol 처리구에서는 무처리 구와 유의차를 관찰할 수 없었다. 한편 모든 ancymidol 관주처리구에서는 초장이 무처리구보다 50~60% 단축 되는 효과를 나타내었다. 그리고 30mg·L−1의 ancymidol 처리구에서는 엽수가 감소하였으며 개화소요일수도 무 처리구보다 다소 지연되었다. Uniconazole 1mg·L−1 처 리구에서 초장을 제외한 모든 형질들은 무처리구와 유 사하였지만 5와 10mg·L−1 처리구에서는 무처리구와 유 의차가 인정되었다. 신나팔나리인 ‘Raizan’ 또한 상기 의 두 품종과 마찬가지로 paclobutrazol 처리구에서는 왜화효과를 관찰할 수 없었다. Ancymidol 처리구는 농도에 관계없이 초장이 무처리구보다 60% 이상 단축 되었으며 특히 30mg·L−1 처리구에서는 개화소요일수가 다소 지연되었다. Uniconazole 10mg·L−1 처리구에서 초장 은 약 26% 감소하였으나 다른 형질은 무처리구와 유의차 가 없었다. 결과적으로, ancymidol과 uniconazole의 관주 처리는 양적 형질에 큰 변화 없이 초장을 줄여 분화용 백합으로 사용하기 위해서 매우 효과적이었다.

진한 붉은색을 띤 ‘Tiny Ghost’를 교배모본으로 Oriental group, Longiflorum group 및 Martagon group을 부본으로 하여 효율적인 종간잡종 생산과 상 품성 있는 신품종을 획득하고자 실험을 수행하였다. 개 화당일 채취한 화분의 발아율은 Oriental hybrid인 ‘Sorbonne’이 64%로 가장 높았다. 주두수분법과 화주 절단수분법을 이용하여 수분한 결과, ‘Aktiva’를 부본 으로 사용한 경우를 제외하고 모두 주두수분법을 사용 한 교배에서 자방이 생성되었다. 또한 Longiflorum group 인 ‘Norina’와 ‘Gelria’ 중 화분발아율이 높은 ‘Norina’ 가 모두 수정에 실패하여 같은 group 내에서도 종간 의 유전적 친화성에 따라 수정률이 달랐다. ‘Aktiva’를 부본으로 한 교배에서 1개의 embryo sac을 적출하여 1개 의 구를 생성하였으며, ‘Sorbonne’을 부본으로 한 교배 에서는 4개의 ovule을 치상하여 1개의 구를 얻었다. ‘Gelria’를 부본으로 한 교배에서는 1개의 자방에서 embryo 5개, embryo sac 15개, ovule 7개를 얻었으며 그 중 18개체가 발아하여 66.7%의 발아율을 나타내었다. Martagon group인 L. hansonii를 부본으로 한 교배에 서는 5개의 ovule을 얻어 그 중 40%가 발아하였다.

캘러스, 반수체, picloram, 전처리양식을 규명할 목적으로 배양효율에 미치는 화분발육 단계와 온도전처리가 미치는 영향에 대한 실험결과는 다음과 같다. ‘Dreamland’의 감수분열중인 약을 현 미경적방법으로 검경하여 본 결과 화뢰의 길이가 23.0-24.9mm는 tetrad 단계이었고 25.0-26.9mm는 uninucleate 단계, 27.0-28.9mm에서는 binucleate 단계 임을 확인할 수 있었으며, 저온 및 고온 온도처리 결 과는 전체적으로 캘러스 및 배 발생율은 저조하였고 특 이한 점은 발견할 수 없었다. Microgametogenesis 단 계별로 약배양 효율을 조사한 결과, late uninucleateearly binucleate 단계인 23.0-28.0mm에서 17.8% callus가 유도되었고, 식물체 재분화율도 6.7%로 나타 나 적정 배양 stage임을 확인할 수 있었다. 유기된 캘 루스를 picloram과 zeatin을 혼용한 MS 배지에 이식 하여 25oC에서 배양하여 식물체를 얻었다.

xBrassicoraphanus, a new synthetic intergeneric hybrid between Brassica rapa L. ssp. pekinensis and Raphanus sativus L., also locally known as ‘Baemoochae’, is an interesting subject for studying polyploidy and genome plasticity in the family Brassicaceae, but very few genomic and cytogenetic information. Here, we analysed the chromosome complements and pairing of the most fertile lines, BB1 and BB5, using dual-color fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) to check their chromosomal segregation stability. The somatic chromosome complement of B. rapa was confirmed to be 2n=20 (2.8~4.8μm), of R.sativus, 2n=18 (2.0~3.3μm), and of xBrassicoraphanus, 2n=38 (2.2~5.0μm). There were eight, eight, and seventeen metacentric pairs and two, one, and two submetacentric pairs in B. rapa, R. sativus, and xBrassicoraphanus, respectively. Additionally, three, two, and five pairs of 5S rDNA and five, three, and eight pairs of 45S rDNA were observed in B. rapa, R. sativus, and xBrassicoraphanus, respectively. This suggests that both B. rapa (AA) and R. sativus (RR) genomes, particularly the rDNA arrays, co-exist in xBrassicoraphanus (AARR) genome. In meiosis I, nineteen bivalents were most frequent, and GISH analysis showed ten bivalents from the A genome. This study would provide a useful information for further genomic study of xBrassicoraphanus and its improvement as a new promising breeding variety.

Genome sequencing researches for considerable numbers of crops and wild plants are being developed. Cytogenetic researches according to chromosome number and size are essential to confirm and comprehend ploidy level and genome size before genome sequencing project is actually conducted. Cytogenetic researches on six food crop plants were carried out by DAPI staining and fluorescence in situ hybridization (FISH) method. Fagopyrum esculentum Moench showed 2n=2x=16, each chromosome length of 1.42㎛ to 1.77㎛, total chromosome length of 13.31㎛, and karyotypic formula of 2n=8m; Phaseolus angularis W.F. Wight, 2n=2x=22, 2.01㎛ to 3.84㎛, total 28.03㎛, 2n=9m+2sm, Perilla frutescens var. japonica Hara, 2n=2x=40, 1.73㎛ to 2.76㎛, total 44.36㎛, 2n=5m+13sm+2st. Chromosome sizes of the other three species such as, Panicum miliaceum L., 2n=2x=36, total chromosome length of 30.83㎛, Sesamum indicum L., 2n=2x=26, 27.39㎛, lpomoea batatas L., 2n=2x=30, total 33.51㎛ were too small for each chromosome type to be identified and analyzed. The result of FISH analysis using 5S and 45S rDNA probe showed species-specific chromosome locations in the genome. These preliminary analyses were carried out to decide which food crop to prioritize for genome sequencing. This work was supported by the “Cooperative Research Program for Agriculture Science & Technology Development (No.PJ009837), Rural Development Administration, Republic of Korea.

참나리(Lilium lancifolium)는 우리나라 자생나리로 생육이 왕성하고 후자리움에 내성이 있으며 주아를 형성하는 등 많은 장점을 가진다. 이를 자방친으로 하여 Asiatic hybrid "Dreamland"의 상향이고 흑갈색의 반점이 없는 형질을 이입시키기 위해 교배를 수행하였다. 참나리와 "Dreamland"의 화분발아율은 각각 30과 60%이었다. 이들 후대의 개화일자별 화분의 평균 발아율은 25-57%로 양친의 중간에 분포하였다. 개화는