Seung-Keun Jong

Proteome Analysis of Wheat Lemma

We report here for the first time on the construction of proteomes from wheat lemma at the anthesis stage. After transfer of lemma proteins to polyvinylidene difluoride membranes, seventy larger spots were subjected to peptide sequence analysis; the amino acid sequences could be described for forty-eight of these proteins. The result suggested that wheat proteins were less N-terminally blocked compared to rice proteins, which are known to have a much higher ratio of N-terminal blocks. We further analyzed the internal sequences of eight blocked proteins by the Cleveland peptide mapping method. Out of these total 56 amino acid sequences, forty-one could be assigned to the corresponding expressed sequence tags (ESTs). The expression profile of lemma proteins was generally similar to that of leaf, and the majority of identified proteins were related to cellular metabolisms. We analyzed the internal sequences of one protein spot present in lemma, which was not present in leaf.

Pollen-tube behavior and embryo development in interspecific crosses among the genusFagopyrum

Common buckwheat (Fagopyrum esculentum Moench) is an agriculturally and pharmaceutically valuable crop due to its wellbalanced essential amino acids and rutin content. However, global mass production of buckwheat is limited because its genetic self-incompatibility results in low seed sets and poor grain yield. Therefore, this study was conducted to classify the modes of pistil-pollen interaction between species belonging to the genusFagopyrum and to determine the optimal combination of outcrosses for the most successful pollinations. Based on the interaction between pistils and pollen, we classified the modes of pollen tube growth during interspecific crosses ofFagopyrum species into five categories: (i) Highly compatible: normal pollen tube elongation and style penetration within 6~24 hours of pollination, (ii) Slightly compatible: delayed (for 1~6 hours) pollen tube elongation and normal style penetration, (iii) Incompatible type I: pollen tube inhibition at the stigma, (iv) Incompatible type II: pollen tube inhibition at the style, and (v) Incompatible type III: pollen tube inhibition at the stylodium. Based on the observed pollen tube elongation and the following embryo development, highly compatible pollinations were found to be crosses betweenF. esculentum x F. cymosum and betweenF. esculentum (thrum)x F. homotropicum.

Rice Proteomics: A Functional Analysis of the Rice Genome and Applications

In this review, we described the catalogues of the rice proteome which were constructed in our program, and functional characterization of some of these proteins was discussed. Mass-spectrometry is the most prevalent technique to rapidly identify a large number of proteome analysis. However, the conventional Western blotting/sequencing technique has been used in many laboratories. As a first step to efficiently construct protein cata-file in proteome analysis of major cereals, we have analyzed the N-terminal sequences of 100 rice embryo proteins and 70 wheat spike proteins separated by two-dimensional electrophoresis. Edman degradation revealed the N-terminal peptide sequences of only 31 rice proteins and 47 wheat proteins, suggesting that the rest of separated protein sports are N-terminally blocked. To efficiently determine the internal sequence of blocked proteins, we have developed a modified Cleveland peptide mapping method. Using this above method, the internal sequences of all blocked rice proteins(i, e., 69 proteins) were determined. Among these 100 rice proteins, thirty were proteins for which homologous sequence in the rice genome database could be identified. However, the rest of the proteins lacked homologous proteins. This appears to be consistent with the fact that about 45% of total rice cDNA have been deposited in the EMBL database. Also, the major proteins involved in the growth and development of rice can be identified using the proteome approach. Some of these proteins, including a calcium-binding protein that tuned out to be calreticulin, gibberellin-binding protein, which is ribulose-1.5-bisphosphate carboxylase/oxygense active in rice, and leginsulin-binding protein in soybean have functions in the signal transduction pathway. Proteomics is well suited not only to determine interaction between pairs of proteins, but also to identify multisubunit complexes. Currently, a protein-protein interaction database for plant proteins(http://genome.c.kanazawa-u.ac.jp/Y2H)could be a very useful tool for the plant research community. Also, the information thus obtained from the plant proteome would be helpful in predicting the function of the unknown proteins and would be useful be in the plant molecular breeding.