Phenotypic traits (physiological characteristics and somatic incompatibility) and genotypic traits (target region amplification polymorphism TRAP) were used to survey the diversity in Chinese Auricularia auricula systematically, which consisted of 32 main cultivated strains. The 27 important and stable physiological indexes were evaluated; somatic incompatibility test (SIT) reaction was described from three aspects: type, pigment, and intensity while intensity clustering alone revealed the phenotypic diversity among the 32 strains;16 pairs of TRAP primer combinations produced 535 unambiguous and reproducible DNA fragments, of these 524 (97.9%) were polymorphic. The phylogenetic trees were constructed by Unweighted Pair-group Method with Arithmetic Averages (UPGMA), which distributed the test strains into four to six major groups respectively. The principal coordinate analysis (PCO) of the three methods (physiological characteristics, SIT intensity and TRAP) exhibited highly similar clustering patterns, revealing that all the test strains can be divided into three groups considered significantly correlated with geographical regions. Results revealed the occurrence of relatively low diversity of A. auricula in the study. The cultivated strains in the same region are more similar physiologically and some strains are suspected to be synonymous. This study suggests that the value for estimating diversity are represented by TRAP＞SIT reaction intensity＞physiological characteristics in A. auricula, and PCO analysis can provided more effective and visible information than the UPGMA clustering dendrogram. Our finding will facilitate future germplasm resources management and breeding program of A. auricula in China.

본 연구는 신품종 ‘미소’ 느타리버섯에 대한 성양식에서의 불화합성 체계를 이해하고자 수행하였으며 이 실험결과 자실체로부터 채취한 단포자의 교배를 통해 신품종 ‘미소’ 느타리버섯이 자웅이체성이고 4극성 교배양식을 나타내는 것을 확인하였다. 꺽쇠연결이 없는 분리된 담자포자는 단핵균사로 판단되었고, 이러한 단핵균사간 화합, 불화합성 여부는 빠른 생육과 활력있는 발달형태를 통해 어느 정도 육안으로도 확인이 가능하였으며 두 단핵 균사체의 대치배양 후에 형성되는 꺽쇠연결을 현미경으로 관찰하여 명확히 확인되었다. 예외적으로 교배가 화합성인 경우 형성되는 이핵 균사체가 대치배양의 한쪽 가장자리에서만 꺽쇠연결이 발견되는 일방향 이핵화 현상이 관찰되었다. 이 실험결과로부터 4극성 교배에 있어서 A1A2B1B2의 이핵 균사체로부터 발생하는 네 가지 유전자 형태, 즉 A1B1, A1B2, A2B1, 그리고 A2B2가 화합성테스트에 의해 결정되었으며 선발된 나머지의 단핵 균사체의 교배형은 Tester 계통을 이용하였다.

Reproductive incompatibility is an important factor to select a specific biological control agent for successful augmentation of the corresponding endogenous population. An endoparasitoid, Cotesia plutellae (Kurdjumov), is an effective control agent to diamondback moth, Plutella xylostella (L.) and has been known to be classified into two groups in terms of reproductive incompatibility. This study analyzed an Korean population of C. plutellae in terms of morphological characters and mitochondrial DNA marker, which did not match with either of two reproductive incompatibility groups. These results suggest that a Korean population of C. plutellae can be involved in a novel reproductive group. For any augmentation program of C. plutellae in Korea, reproductive incompatibility should be seriously considered to select a particular exotic population.

십자화과 작물에서 SRK와 SP11 유전자는 자가불화합 반응을 매개하는 주요 유전자이다. 무(radish)에서 S-locus haplotype을 분류 및 확인하기 위한 첫 단계로, SRK와 SP11유전자의 온전한 서열확보를 위해 기존 연구를 통해 밝혀진 Brassica rapa의 SRK유전자 서열을 활용해 local blast를수행했다. 이를 통해 무 draft genome sequence에서 SRK유전자와 높은 상동성이 있는 15개의 후보유전자들을 찾았다. 이후 B. rapa genome data를 활용한synteny analysis를 통해 무 draft genome sequence에서 B.rapa의 S-locus region과 synteny를 가지는 scaffold를 R7 연관그룹에서 확인했다. 해당 scaffold에서 SRK와 SLG 유전자의 서열을 확보할 수 있었다. 이렇게 확보한 SRK유전자서열의 정보를 통해 NCBI database에서 동일한 유전자서열을 찾을 수 있었고, 해당 논문에서 연구된 같은 haplotype의 SP11유전자서열을local blast의query로 사용해서 무 draft genome sequence에서 SP11 유전자정보가 포함된 scaffold를 찾을 수 있었다. 이로써, SRK, SLG, SP11/SCR 유전자를 포함하는 53,785bp, 42,804bp, 10,165bp 크기의 온전한 genomic 서열을 확보하게 되었다. 무 S locus haplotype을 분류하기 위한 체계를 만들기 위해 S-locus core region에 있는 SLL2유전자를 활용했다. SRK 유전자의 경우, 무 genome내에 상동성이 높은 homologous gene을 가지고 있고, SP11유전자의 경우는 exon지역의 다형성이 너무 높아PCR기반의 marker개발이 어렵기 때문에 SLL2유전자를 활용했다. 가지고 있는 다양한 무 육종계통에서 SLL2 유전자에 특이적인 primer set을 사용해 SLL2유전자를 증폭시킨 뒤, sequencing하여 SLL2유전자에 대한 다양한 대립유전자들의 서열을 확보할 수 있었다. 확보한 SLL2대립유전자 서열을 비교함으로써 S locus haplotype분류체계를 만드는데 활용 가능한 conserved region을 exon2와 exon6에서 확인할 수 있었고, 해당 부분에 design된 primer를 통해 다양한 무 육종계통에서 단일한 PCR band를 확인할 수 있었다. 이는 직접적인 sequencing을 통해 S locus haplotype을 식별하는데 충분한 정보를 제공함으로써 무 육종에 큰 도움이 될 것이라 생각된다.

사과는 배우체형 자가불화합성을 나타내는데 이는 S-locus의 복대립유전자에 의해 조절된다. 본 연구는 S-allele specific PCR분석을 통해 신품종을 포함한 24종의 사과 주요 재배품종과 7종의 꽃사과 품종의 자가불화합성 유전자형(S-genotype)을 결정하고자 수행하였다. 31종의 재배품종과 꽃사과 품종 을 23종의 S-allele specific primer을 이용하여 분석한 결과 12개의 S-allele (S1, S2, S3, S5, S7, S9, S10, S16, S21, S23, S26, S29)이 동정되었다. 그 중에서 24종의 재배품종에는 S1(41.7%), S3(58.3%), S7(29.2%), S9(54.2%)의 S-allele이 흔히 존재하는 것으로 확인되었다. 국내육성 신품종인 ‘아리수’와 ‘황옥’의 S-genotype은 각각 S3S7과 S3S9으로 동정되었다. 본 실험에서 얻은 S-genotype 정보는 안정적인 사과 과실생산에 적합한 수분수 선발과 육종프로그램에서 교배조합 작성에 유용 하게 활용될 것이다.

The different forms of flowers in a species have drawn thoughtful attention of many evolutionary botanists, including Charles Darwin. Common buckwheat (Fagopyrum esculentum Moench.) is regarded as a dimorphic self-incompatible plant which bears either a pin or a thrum flower. It is revealed that the S supergene the key element to govern the self-incompatibility, flower morphology, and pollen size. Already, we have produced self-incompatible buckwheat lines by an interspecific cross between F. esculentum and F. homotropicum by using embryo rescue. We also notice that the self-compatibility allele, Sh, keeps up the heteromorphic incompatibility. In the past decades, two dimensional gel electrophoresis based proteomics approaches have been applied systematically to identify and profile proteins expressed during pollen development of model plant species. Proteome techniques have vastly been applied in the fields of plant genetics, plant development, and plant physiology and ecology to reveal plant genetic diversity, plant development, differentiation of plant tissue and organ, separation and functional identification of novel component of various organelles, mechanisms of plant adapted to abiotic or biotic stresses including high temperature, low temperature, high salt, drought, and pathogens and insects, and interaction of plant with microbe. However, the plethora of studies related to heteromorphic has added remarkably to our knowledge in the field of the multiple aspects of the breeding system and many researches have provided evidence for the connection between these two components. But in spite of its potential biological significance, the high throughput proteomics analysis of this connection has so far been grossly overlooked. So our attempts are to unravel the proteome investigation in common buckwheat.

Common buckwheat has the sporophytic self-incompatibility mechanism and that’s why it has the ability to cross pollinate between two plants with different styles (thepin type and thrum type). The S supergene is thought to govern self-incompatibility, flower morphology and pollen size in buckwheat. Already, we have produced self-compatible buckwheat lines by an interspecific hybridization between Fagopyrum esculentum and F. homotropicum by embryo culture. The pollen size of F1 plants produced by a cross between a pin type plant and the self-compatible plant was similar to that of the self-compatible lines and segregated together with flower morphology without exception. The pollen tubes of the self-compatible plants were compatible with styles of the pin plants but incompatible with the styles of thrum plants. But, the pollen tubes of thrum flowers were compatible with the styles of self-compatible plants. Also, the pollen tubes of pin flowers were incompatible with the styles of self-compatible plants. Already, from these results, we have reported a tentative genotype for heterostyle and homostyle flower types. Homomorphism was controlled by a single allele Sh, while the pin/thrum-complex gene was governed by a single genetic locus S, with two alleles, S and s, which control Ss (thrum-type) as well as the ss (pin-type), respectively. Corresponding represents the case of a single locus S with three alleles, Sh, S and s, and the phenotypes, homomorphic, pin and thrum. It can be characterized by relationship of dominance, S>Sh>s. Using the two self-fertile lines, one is considered as the long-homostyle flowers and the other is considered as the short homostyle flowers. If the short-homostyle trait had arisen by recombination in the S supergene, its genotype would be considered to be GIs ip a/GIs ip a. The pollen tubes of the short-homostylous plant should be compatible with the styles of thrum plants. Also, the pollen tubes of short-homostylous plants should be incompatible with the style of long-homostylous plants, and the reciprocal cross also should be incompatible, because the genotype of long homostyle is gis Ip PA/gis Ip PA. Furthermore, the flower morphology of F1plants produced by the cross between cross and short homostyle flowers should be thrum or short homostyle and only short-homostylous plants should be produced by the cross between pin and short homostyle flowers. However, the compatibility or incompatibility of short homostyle flower was not clarified. So, we need to clarify the compatibility or incompatibility of the style of short homostyle flowers for the next step.

본 연구는 사과의 자가불화합성 현상에 효과적인 저해제와 활력제를 탐색하기 위해 여러 종류의 물질을 조사하였다. 후지 사과의 화주조직에서 분비되는 S RNase에 대한 저해효과를 조사하기 위해 In vitro 상에서 효소 저해제를 넣어 효소활성을 측정한 결과, Iron(II) Sulfate에서 현저하게 활성이 24.3% 감소되어 저해효과가 뛰어난 것으로 나타났다. In vitro 상에서 후지 사과의 화분관 신장 억제 효과를 분석한 결과, 후지 사과의 화분은 2% Sucrose 신장배지 보다 10% Sucrose 신장배지에서 약 90% 이상의 높은 발아율을 나타났다. 또한 저해제와 첨가제 처리에 의한 안동지역과 영주지역의 Field에서 후지 사과 착과율 조사 결과, 안동지역에서는 A++V6 2회 처리가 86.1%로 중심화착과율이 가장 높았으며, 영주지역에서는 A++V1 1회 처리가 91.9%로 중심화 착과율이 가장 높은 것으로 나타났다.

Apricot (Prunus armeniaca L.) cultivars show a gametophytic self-incompatibility (GSI) system, like other fruit species of Rosaceae family. Thus, it is necessary to determine their S-genotypes in order for stable fruit set in commercial cultivation. S-genotypes of apricots were determined by PCR and test crosses. Three sets of consensus primers designed from Prunus S-RNases were used to amplify fragments containing the first and second S-RNase intron, respectively. Through the results obtained from the 3 PCRs, we could identify SI genotypes of 33apricot cultivars. Several cultivars such as 'Heiwa', 'Yamagata No.3' and 'Shinsuoomi' had the self-compatible (Sc) allele. Self-pollination tests revealed that cultivars with Sc allele were self-compatible. Cross-pollination tests confirmed that there was cross-incompatibility between the cultivars with the same S-genotypes. These results might be very useful for growers for effective pollination and for breeders using these in cross breeding programs.

Self-incompatibility (SI) prevents self-fertilization by inhibiting the pollen tube growth of self-pollen. Molecular analysis has revealed that the S locus comprises a number of genes, such as the S-locus glycoprotein (SLG), the S-locus receptor kinase (SRK), and SP11 (SCR). Although molecular markers related to those genes have been developed, a simple S-haplotype detecting method has not been reported due to the highly polymorphic and relatively small coding regions. In this study, the sequence characterized amplified region (SCAR) markers were used to establish an efficient radish genotyping method. We identified the S-haplotypes of 192 radish accessions using 19 different markers, which proved to be highly reliable. The accessions were assigned to 17 types of S-haplotypes, including 8 types of SRKs and 9 types of SLGs. Since the developed SCAR markers are based on their gene sequences, we could easily identify the S-haplotypes by a single specific band, with the highest frequencies detected for SLG 5, SRK 1, and SLG 1, in order. Among the tested markers, the SLG 1, SRK 1, and SRK 5 markers exhibited high reliability, compared to phenotypic results. Furthermore, we identified the seven types of unreported SLGs using SLG Class -I and -II specific markers. Although the developed SCAR markers still need to be improved for the genotyping of all S-haplotypes, these markers could be helpful for monitoring inbred lines, and for developing the MAS in radish breeding programs.

황기의 자가불화합성 타파를 위한 노화수분 처리, NaCl 처리 및 CO2 처리에 대한 결과를 요약하면 다음과 같다. 비닐하우스에서 노화수분은 8월 하순에는 거의 수정이 되지 않았으며, 9월 상순부터 10월 상순까지는 개화 당일 부터 개화 후 2일까지 수정되었고, 9월 하순 개화 당일 수분한 것이 결협율 33.3%, 결실율 86.2%로 가장 양호하였다. 노지에서 노화수분은 대조구는 8월 하순부터 10월 상순까지 결협은 되었으나, 9월 중순과 9월 하순을 제외하고 결실이 되지 않았으며, 9월 하순의 개화 후 1일에 결협율 39%, 결실율 94.9%로 가장 양호하였다. NaCl 처리는 비닐하우스에서는 1% 농도 처리가 결협율은 10월에서 21.3%로 가장 좋았으나, 결실율은 9월 하순 66.7%로 가장 양호 하였고, 노지에서는 NaCl 농도 5%에서 결협율 7.3%, 결실율 90.9%로 가장 양호하였다. 식물생육상을 이용하여 CO2 700 ppm을 처리한 결과 무처리구와 큰 차이 없었다. 결론적으로 황기 자가불화합성 타파를 위하여 9월 중하순에 자화가 아닌 자가화를 개화당일 또는 개화 후 1일에 인공수분해주거나 NaCl 1~5%를 처리해주면 자가불화합성을 타파할 수 있다고 사료된다.

This study was conducted to determine the characteristics of fertilization process and embryo development of Astragalus membranaceus Bunge (Astragali Radix) to provide basic data needed in its breeding. A. membranaceus showed poor seed setting when self-pollination was induced. When artificial pollination was induced, it showed less than 5% bearing in late August, but more than 13% bearing from the beginning of September 4th. The flower size was about 17.0 mm×4.0 mm and pistils and stamens had the same length of 15.0mm at flowering stage. When self-pollination or cross-pollination was induced, pollen tubes extended to an ovule. While pollen tube was extending to the ovule, reproductive cell split and formed two male generative nuclei and a vegetative nucleus. In the case of self-pollination, fertilized embryo was not observed, but was formed in the case of cross-pollination. A. membranaceus is noted to have zygote self-incompatibility. In the case of cross-pollination, fertilization was observed in 6 to 8 h after pollination, where apical cell derivatives split after fertilization. A spherical pro-embryo was then formed three days after fertilization. The seed attained full shape with a seed coat showing its distinctive contour 15 days after fertilization. Thus, A. membranaceus in Leguminosae family is found to have zygote selfincompatibility although its flower shape is shown to match the self-compatibility plant.

사과 재배시 대두되고 있는 수정 불량 및 안정된 수정을 통한 품질 향상을 위한 방안을 모색하고자 자가불화합성 제어제(상품명 : 애플플러스)를 이용한 SSR 유전 분석을 통한 자가수정 향상 정도 및 촉진 효과, 품질 특성에 미치는 영향을 검토하였다. 1. 후지 품종에 있어서 중심화 결실율은 자연방임시 무처리의 경우 2004년 0.5%, 2005년 3.0%인데 비하여 자가불화합성 제어제 처리의 경우 2004년 17.2%, 2005년 44.4%로 높게 나타났다. 2. 망을 씌워 자가불화합성 제어제를 처리한 실험구에서도 무처리구에서는 2004년 0.4%, 2005년 0.7%, 처리구에서는 2004년 3.3%, 2005년 5.2%의 결실율을 보여 처리구가 7.4~8.3 배정도 결실율이 높게 나타났다. 3. 망을 씌워 자가불화합성 제어제 처리한 실험구와 무처리구를 비교하면 L/D, 경도에서는 유의성이 인정되지 않았으며, 종자수에서는 무처리구 0.9개, IS-1 처리구 1.2개, IS-2 처리구 1.2개, IS-3 처리구 1.5개, IS-4 처리구 0.7개로 나타나 약제 종류에 따라 효과가 다르게 나타났다. 4. SSR 유전자 분석에 의한 후지 품종 종자의 자가수분 정도는 무처리구에서 0%, IS-1 처리구 100%, IS-2 처리구 73%, IS-3 처리구 88%, IS-4 처리구 62%로 나타나 IS-1의 약제가 가장 높게 나타났다.

Korean sweetpotato, Ipomoea batatas, varieties were examined for the cross-compatibility and -incompatibility through inter-crosses among them to obtain the fundamental genetic informations for enhancing the heterosis breeding efficiency. The pollen germi