Selection Indexes and Genetic Advances in Soygbeans and their Implications to Selection

CHANG, KWON YAWL,HAN, KYUNG SOO,KO, MI SUK,KIM, JIN HYEONG 경상대학교 유전공학연구소 1986 遺傳工學硏究所報 Vol.5 No.-

This experiment was conducted to estimate the selection index and genetic advance for nine quantitative characters of soybeans. F_1's and F_2's were planted in a randomized block design with three replications, and the data were analyzed by the methods of Robinson et al. 'b' value for stem diameter was the highest among the characters, and for days to flowering was positive in F_1 generation, but negative in F_2 generation. The highest genetic advance value was expressed as the combination of all characters but in spite of the expected usefulness of selection index techniques, unreasonable problems such as the expence, time and labor were remained. For these reasons, it was realized that the selection index should be calculated on a basis of the data of three characters, i.e., days to flowering, stem diameter and pod numbers per plant.

The Role of the Extracellular Cellulase Gene in Xanthomonas oryzae pv.oryzae on Bacterial Leaf Blight

YUN, HAN DAE,PARK, YOUNG WOO,LIM, SUN TECH,KIM, HEE KYU,KANG, KYU YOUNG 경상대학교 유전공학연구소 1992 遺傳工學硏究所報 Vol.11 No.-

A plasmid, designated pXC3-43, encoding an extracellular cellulase activity from Xanthomonas oryzae pv. oryzae(Ishima) was reported previously by authors^1). The cellular distribution of the endoglucanase activities of pXC3-43 and X. o. pv. oryzae was almost exculusively extracellular. Total endoglucanase activity expressed by the pXC3-43 was about 80% of X. o. pv. oryzae k1 wild type. Tn5 mutagenesis of pXC3-43 revealed that extracellular cellulase structural gene is located in a ca. 5 kb KpnI fragment. To examine the role of the extracellular cellulase in disease process, three cellulase-minus mutants of X. o. pv. oryzae were constructed by marker exchange of negative Tn5 insertions into the chromosome of the wild type. These cellulase -minus mutants were as virulent as the wild type in pathogenicity test on leaves of rice(Oryzae sativa L.). Furthermore, When investigated under transmission electron microscopy (TEM), bacterial leaf blight(BLB) symptoms by the cellulase-minus mutants, X. o. pv. oryzae K1::Tn5 were similar to those by wild type. These results suggest that the extracellular cellulase of X. o. pv. oryzae is not primary mechanism of pathogenesis for the pathogenesis of BLB, but mightbe involved in an early disease developmental stage.

Reconstitution of Artificially Designed inaz Gene and Its Expression in vivo

BAHK, JEONG DONG,CHO, MOO JE,LEE, DAE SIL 경상대학교 유전공학연구소 1992 遺傳工學硏究所報 Vol.11 No.-

In order to express the artifically designed inaz gene, which was deduced from the InaZ protein of Pseudomonas syringae, in Escherichia coli, a fused protein was planned. One unit of the fused protein consists of N-term. 50 amino acids of LacZ protein of E. coli and one unit of the repeated 48 amino acids from the core domain of InaZ protein of Ps. syringae. Each of them consists of eight synthetic DNA fragments. All of them were phosphorylated, block-ligated and reconstituted. Theresfter, the reconstituted DNA stretah was inserted into the plasmid vector pASI containing a P_L promoter of bacteriophage lambda. The recombinant plasmid pASLF was transformed to an expression host, E. coli N4839-1. After performing expression under a permissible or impermissible temperature, the cultures were applied to the buffer gradient gel and ckecked. At the near of 12-13 kD region a clear thick band was observed to be the fused INA protein.

Characterization of A New Chromosomal Gene Which Maskes dsRNA Yirus Cytopathology in ski^- Saccharomyces cerevisiae

HWANG, BO HYE,SEO, SOOK JAE,LEE, HYUN SOOK 경상대학교 유전공학연구소 1991 遺傳工學硏究所報 Vol.10 No.-

The yeast L-A virus(4.6 Kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and M₁, RNA satellite virus a 1.8 kb dsRNA encode a secreted protein toxin(the killer toxin). The yest chromosomal SKI genes, so named for the superkiller phenotype of mutants, prevent viral cytopathology by lowering the virus copy number. Thus, ski^-mutants are either temperature sensitivity(ts) or cold sensitivity(cs) for growth. Under electron microscope virus-infested ski^- mutants showed abonormal budding pattern. We transformed a ski2 virus infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant recovered from typical ski^- cold sensitivity though it remained superkiller. Elimination of the DNA clone from the ski2 strain eliminated this phenotype and introduction of the VPMs(Viral Pathology masking gene in Ski^- strain) DNA recovered from such transformants into the parent ski 2 strain or into ski3 or ski6 mutants gave the same phenotype. These transformants recovered from abnormal budding pattern. Compare of dsRNA contents in VPMs transfomed cell with control cell transformed only YEp13 vector, dsRNA transformed cells were not changed. Thus VPMs is a noe chromosomal gene which maskes dsRNA virus cytopathology in ski^- background. The 10 kb insert showed this activity in ski^- virus infested cells.

Molecular Cloning of Genetic Determinants of Pectate Lyases from Rhizobium fredii USDA 193

YUN, HAN DAE 경상대학교 유전공학연구소 1990 遺傳工學硏究所報 Vol.9 No.-

Rhizobium fredii USDA193 is a microsymbiont of Glycine max(soybeans) which was originally isolated from People's Republic of China^19). It infects and nodulates soybean roots via hairs by actively penetrating the root hair cell-wall. The penetration of Rhizobium into the root hair cell-wall requires the activity of cell-wall hydrolyzing enzymes such as cellulases and pectinases. Evidence is presented that R. fredii USDA193 produces pectate lyase(pel) enzymes and that their genetic determinants are borne on the resident symbiotic plasmid(pSym) DNA. The pectate lyase genes pelB and pelE of Erwinia chrysanthemi EC16 show homology to a 4.0 Kb EcoR1 fragment of pSym DNA. The pelB and pelE homologues in R. fredii USDA193 are tandemly located on a 0.7 Kb Sal1-HindⅢ and 1.3 Kb HindⅢ- EcoR1 fragment, respectively. These two subfragments carrying the homologous RpelB and RpelE(for Rhizobium pel genes) genes, have been subcloned into high-express pectate lyase enzyme activity in an Escherichia coli background.

Subcloning and Sequencing of β-Xylosidase Gene of Alkalophilic Bacillus sp. K17

SUNG, NACK KIE,SHIM, KI HWAN,CHUN, HYO GON,CHUNG, CHUNG HEE,CHUNG, YOUNG CHUL,KO, HAK RYONG 경상대학교 유전공학연구소 1988 遺傳工學硏究所報 Vol.7 No.-

The 5.0kb insert of pAX278, which was previously cloned, was reduced by various deletion and thus I. 4kb EcoRI-XbaI fragment was recloned into pUC19(pAK208). To increase β-xylosidase activity, the I. 4kb insert in pAK208 was recloned into expression vector, pKK223-3 (tac-promoter), and β-xylosidase activity in cell carring this plasmid (pKHR212) was increased was 3.5 times than that in pAX278. The complete nucleotide sequence of thermophilic alkalopilic Bacillus sp. K17 and its flanking regions were established. A 1263-bp open reading frame for β-xylosidase was observed. The molecular weight(50, 521 dalton) deduced from for β-xylosidase gene, was agreed with the result obtained by SDS-polyacry-lamide gel electrophoresis of the purified enzyme. The Shine-Dalgarno sequence was found-8 bp upstream of the initiation codon ATG. The 1-10 regions for promotor sequence of β-xylosidase gene were similar to the consensus sequence for B. subtilis RNA polymerase, and the-35 regions were different from all the known promotor for B. subtilis RNA polymerase.

Genetic Stability of the Characters Transformed by Microinjection of Exogenous DNA into Crop Plants

KANG, NAM-JUN,PARK, CHAN-HEE,JEONG, YEON-OK,CHO, JEOUNG-LAI,KIM, ZHOO-HYEON,UM, SUNG-KYUN,PARK, JUNG-CHOON,KANG, SEONG-MO 경상대학교 유전공학연구소 1988 遺傳工學硏究所報 Vol.7 No.-

Genetic stability was examined for the second generation(TS-2) progenies of 4 transformants induced by injecting Altari(A) DNA into kungzung(K) radish. All TS-2 plants originated from T3 and T4 had red hypocotyls, suggesting the stable incorporation of the foreign gene and its expression at the second generation. The progenies of these 2 transformants had bigger leaves and heavier roots. However, the segregation of hypocotyl colors of TS-2 plants from T5 and T7 was variable, although the majority still had red hypocotyls. Except the progenises derived from T7, the shapes of cotyledons and the first true leaf, and overall morphology of TS-2 plants as well, were of A types as long as their hypocotyl color was red. For the seedlings to have red hypocotyls, the banding patterns of total suluble proteins and the isozymes of MDH and acid phosphatase should be similar to those of A radish, or at least a mixed type of A and K radish. The later the embryos were removed from the mother plants, and the higher the concn of exogenous DNA up to 10㎍ embryo, the better the differentiation of the explants. The probability to porduce intergeneric hybrids between radish and Chinese cabbage (CC) depended on which was being served as a female. The hybrid plants tended to have mixed banding patterns of total proteins and the isozyme when CC served as a female

Induction of Genetic Transformation Through Protoplast Fusion and Microinjection of Foreign DNA into the Ovaries or Embryoids.

PARK, CHAN HEE,KANG,NAM JUN,UM, SUNG KYUN,KANG, SEONG MO,CHO, JEOUNG LAI,KIM, ZHOO HYEON,PARK, JOONG CHOON 경상대학교 유전공학연구소 1989 遺傳工學硏究所報 Vol.8 No.-

Tomato protoplasts were cultured to microcalli in an agarose system. The plating efficiency was high in Ailsa Craig and Mill, and low in Red Cherry. The development of an agarose culture method reduced the amount of work needed to change the culture medium involved in conventional liquid culture system. Tomato protoplasts isolated from two different species were fused with polyethylene glycol, and calli were obtained from the fused protoplasts. A successful cell division and colony formation was obtained from the protoplasts isolated from tomatoes and co-cultivated with the DNA isolated from potato leaves. Using a liquid culture medium combined with filter paper wicks effective in reducing the contamination problem to culture pepper anthers. The highest(41%) callus formation was obtained by using the Sibi et al medium, but only when supplemented the medium with 10 ppm IAA, 5 ppm 2,4-D and 10 ppm kinetin. Among the progenies from radish transformants initially obtained by microinjection of foreign DNA, the test fro the genetic stability of the expression of hypocotyl color indicated that all forty progenies from one transformant were of DNA donor type up to the third generation. It is conclueded that the microinjection of foreign DNA could be used to induce genetic transformation of crop plants.

Sequence structure and overexpression of a cloned β-glucosidase gene from Pseudomonas sp. LBC505

KIM, YANG WOO,CHUN, SUNG SIK,CHUNG, YOUNG CHUL,SHIM, KI HWAN,SUNG, NACK KIE 경상대학교 유전공학연구소 1992 遺傳工學硏究所報 Vol.11 No.-

The nucleotide sequence of the β-glucosidase gene, coding for the β-glucosidase of Pseudomonas sp. LBC505 was determined. An open reading frame identified could encode a protein of Mr 38,5000, which is close to the size(about 40,000) of the polypeptide determined by SDS-gel electrophoresis of crude enzyme. Analysis of the amino-terminal residues of the protein produced in Escherichia coli suggested that it had the signal peptide processed by a signal peptidase. A sequences within the upstream of the β-glucosidase gene were found act as a promoter and a Shine-Dalgarno(SD) sequence for expression vectors, pKK223-3 and pASI, which were regulated by IPTG and temperature respectively. Under the maximum conditions the recombinant β-glucosidase genes, pKKB10 and pASB20, were expressed 8 and 6.2-fold higher levels of β-glucosidase activity than pGL 1, respectively.

GENETIC RELATIONSHIPS BETWEEN AMINO ACIDS, FATTY ACIDS, AND SOME CHARACTERISTICS OF SOYBEAN SEEDS BY THE DIALLEL CROSSES

YANG, MIN SUK,YUN, HAN DAE 경상대학교 유전공학연구소 1983 遺傳工學硏究所報 Vol.2 No.-

The electrophoretic patterns of proteins, amino acid, fatty acid compositions, and minerals in soybean seeds cropped from six parents and 15 diallel crossed F_1 hybrid were analyzed in order to examine gene distributions, combining ability and the relationship between parents and 15 F_1 hybrids. Seven essential amino acids, namely threonine, valine, methoinine, isoleucine, leucine, phenylalanine, lysine, were identified and F_1 generation of this diallel set of crosses was analysed by the method of Griffing and Hayman. Protein and lipid contents of soybean seeds were found to be varietal characteristics, and the results showed soybean plant producing high protein showed low lipid content. The major fatty acid identified was linoleic adid, which acted inversely compared to other fatty acids A total of 25 to 27protein bands were identified from all samples by SDS-polyacrylamide gel electrophoresis. Varietal difference, in mobility of the band were not observed in all samples, but three bands, ^#6, ^#7 and ^#20, were distinguished in each sample. In accordance with the results of 6 parents, the differences in patterns of band^#6, ^#7 and ^#20, were identified in 15 F-1 hybrid. Form the results of amino acid composition and its genetic relationship by statistical analysis, dominance of variance by heterosis was determined to be larger than the additive component of variance, and over dominance was expressed for all essential amino acids. Mean squares of GCA was greater than those of SCA for threonine, methionine and lysine. The effects of GCA for amino acids were different from those of parents and the effects of SCA for amino acids were also different from those of parents.