• The Korean Journal of Mycology
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• v.50 no.4
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• pp.281-289
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• 2022

Fusarium head blight is an important disease of small grains. It is mainly caused by members of the Fusarium graminearum species complex (FGSC). Barley and wheat growers spray fungicides, especially demethylation-inhibitor fungicides, to suppress the disease. The objective of this study was to examine the changes in the sensitivity of the FGSC population to the triazole fungicide, propiconazole. A total of 124 and 350 isolates of FGSC were obtained from barley and wheat in Jeolla Province during 2010-2016 and 2020-2021, respectively. The species identity and trichothecene chemotypes of the FGSC isolates were determined based on polymerase chain reaction assays targeting translation elongation factor 1-alpha and TRI12 genes, respectively. Sensitivity to propiconazole was determined based on the effective concentration that reduced 50% of the mycelial growth (EC₅₀) using the agar dilution method. Of all isolates, F. asiaticum with the nivalenol chemotype was the most common (83.9% in 2010-2016 and 96.0% in 2020-2021), followed by F. asiaticum with the 3-acetyl deoxynivalenol chemotype (12.1% in 2010-2016 and 2.9% in 2020-2021). The EC₅₀ values of the isolates collected in 2010-2016 and 2020-2021 ranged from 0.0180 to 11.0166 ㎍/mL and 1.3104 to 17.9587 ㎍/mL, respectively. The mean EC₅₀ value of the isolates increased from 3.8648 ㎍/mL in 2010-2016 to 5.9635 ㎍/mL in 2020-2021. The baseline resistance to propiconazole was determined to be 7 ㎍/mL, based on the EC₅₀ value of isolates collected in 2010-2016, and the ratio of resistant isolates increased from 9.7% in 2010-2016 to 28.6% in 2020-2021.

• Korean Journal of Ichthyology
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• v.35 no.2
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• pp.91-100
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• 2023

The early life history of Silurus microdorsalis living in Jahocheon Stream was studied by observing egg and morphological development. Live fish were captured in June 2018, then reared in a circulating filtration system under a 14L : 10D photoperiod with a water temperature of 18℃. To artificially induce spawning, females were injected with 0.5 mL of Ovaprim (Syndel, Nanaimo, BC, Canada) per kg of body weight, and males were injected with 10,000 IU/kg body weight of human chorionic gonadotropin. Approximately 15 h later, eggs were artificially inseminated by the dry method. Mature eggs were light pale yellow, which separated them from immature eggs. Fertilized eggs were 2.16±0.06 mm (n=8) in diameter and fully hatched at 181 h after fertilization. The fertilization rate was 63.1±2.2%, and 10.0±3.7% of the embryos were malformed at 18℃. The rates of development were 181 h at 18℃, 109 h at 21℃, and 76 h at 24℃. The larval size immediately after hatching was 4.64±0.22 mm (n=8), and the larvae displayed negative phototaxis at 1 day after hatching. The total larval length on 7 days after hatching was 12.47±0.53 mm, with 25~30 basal anal fin rays and 14~16 basal caudal fin rays observed. The total larval length was 14.13±0.51 mm on 9 days after hatching, and approximately 90% of the black endoplasmic reticulum was deposited on the head and body. The dorsal fin had formed, and a single basal body was observed. On 15 days after hatching, the total larval length was 16.69±0.31 mm; the number of basal caudal fin rays (18 poles) was an integer because 2 dorsal fin basal rays and 60~63 anal fin basal rays were observed. The total larval length was 28.96±1.10 mm on 50 days after hatching; the numbers of caudal fins (n=18), dorsal fins (n=3), pectoral fins (n=11), and anal fin basal rays (n=67~73) were integers.

• Journal of the Korean Society of Food Culture
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• v.21 no.3
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• pp.297-302
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• 2006

This study has examined the physicochemical properties of the Pleurotus eryngii, including their proximate components, amount mineral content, total dietary fiber, total sugar, reducing sugar and free sugar. Additionally, it measured the P. egii ethanol extracts and the total amounts of polyphenol compounds as well as its electron donating ability (EDA) of the substance fraction (SF). The P. eryngii powder's moisture content was 9.0% and each of the other element content such as carbohydrate, crude protein, crude ash and crude fat was found to be 63.06%, 20.70%, 5.20% and 2.0% respectively. Potassium (K) was shown to be the greatest inorganic content and manganese (Mn) was the lowest. Furthermore fructose, galactose, glucose lactose and maltose free sugar content was found in this order. 387 mg% of the total amounts of polyphenol was found in the P. eryngii whole body ethanol extract, 158 mg% in the stipe extract, 593 mg% in the pileus extract and 607 mg% in the substance fraction (SF). Electron donating ability (EDA) was highest at 91.12% in the whole body extract and lowest at 62.90% in the stipe extract. Additionally, the EDA for substance fraction (SF) 0.02%-0.1% was found to be 57.78-77.33%, which was lower than the 0.02%-tocopherol (93.92%) and BHT (96.72%). From these results, it can be assumed that P. eryngii offers superior antioxidative effects with its high content of fiber, inorganic materials and total amounts of polyphenol as well as high electron donating ability (EDA), thereby making it ideal for use in functional foods and industrial materials.

• Journal of Life Science
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• v.33 no.10
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• pp.828-834
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• 2023

The cDNA that encodes transmembrane protein 258 (Tmem258) was cloned from Gryllus bimaculatus and named GbTmem258. This protein comprises 80 amino acids, has no N-glycosylation site, and contains five potential phosphorylation sites at two serines, two threonines, and one tyrosine. The predicted molecular mass of GbTmem258 is 9.06 kDa, and its theoretical isoelectric point is 5.5. The tertiary structure of GbTmem258 was predicted using the available secondary structure information, which suggests the presence of alpha helices (52.5%), random coils (22.5%), extended strands (16.25%), and beta turns (8.75%). Homology analysis revealed that GbTmem258 exhibits high similarity at the amino-acid level to Tmem258 found in other species. The effect of starvation and refeeding on GbTmem258 mRNA expression was also examined in this study. It was found that GbTmem258 mRNA expression in the hindgut progressively increased throughout the starvation period, peaking at almost 1.5 times the control level after six days of starvation. However, refeeding for one to two days after the six-day starvation period restored GbTmem258 mRNA expression to the control level. In fat body, GbTmem258 mRNA expression was almost 3-fold higher during starvation compared to the control level. Refeeding for one to two days after the six-day fast resulted in a decline in the expression to about a 2.5-fold increase over the control level. Throughout the starving and refeeding periods, no other tissues showed any discernible alterations in GbTmem258 mRNA expression.

• Journal of Life Science
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• v.33 no.10
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• pp.808-819
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• 2023

This study was conducted to develop a selection marker for the identification of the Bacillus licheniformis K12 strain in microbial communities. The strain not only demonstrates good growth at moderate temperatures but also contains enzymes that catalyze the decomposition of various polymer materials, such as proteases, amylases, cellulases, lipases, and xylanases. To identify molecular markers appropriate for use in a microbial community, a search was conducted to identify variable gene regions that show considerable genetic mutations, such as recombinase, integration, and transposase sites, as well as phase-related genes. As a result, five areas were identified that have potential as selection markers. The candidate markers were two recombinase sites (BLK1 and BLK2), two integration sites (BLK3 and BLK4), and one phase-related site (BLK5). A PCR analysis performed with different Bacillus species (e.g., B. licheniformis, Bacillus velezensis, Bacillus subtilis, and Bacillus cereus) confirmed that PCR products appeared at specific locations in B. licheniformis: BLK1 in recombinase, BLK2 in recombinase family protein, and BLK3 and BLK4 as site-specific integrations. In addition, BLK1 and BLK3 were identified as good candidate markers via a PCR analysis performed on subspecies of standard B. licheniformis strains. Therefore, the findings suggest that BLK1 can be used as a selection marker for B. licheniformis species and subspecies in the microbiome.

• Journal of Food Hygiene and Safety
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• v.38 no.5
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• pp.319-331
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• 2023

Aminoglycoside antibiotics, also known as aminoglycosides (AGs), are veterinary drugs effective against a wide range of gram-negative and gram-positive bacteria. Owing to their recent use in cultured meats, it has become essential to establish an analytical method for safety management. AGs are highly polar compounds, and ion-pair reagents (IPRs) are used to ensure component separation. Owing to the high possibility of potential mechanical problems resulting from IPR addition to the mobile phase, an analytical method in which IPRs are added directly to the vial was explored. In this study, methods for analyzing 10 AGs via liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the addition of two IPRs were validated for selectivity, detection limit, quantitation limit, recovery, and precision. The detection limit was 0.0001-0.0038 mg/kg, the quantification limit was 0.004-0.011 mg/kg, and the linearity (R²) within the concentration range of 0.01-0.5 mg/kg was over 0.99. Recovery and precision (expressed as relative standard deviation) evaluated in the two matrices (beef and cell culture media) ranged from 70.7% to 120.6% and 0.2% to 24.7%, respectively. The validated AG analytical method was then applied to 15 meats prepared from chicken, beef, and pork, and 6 culture media and additives used in cultured meat. No AGs were detected in any of the 15 meats distributed in Korea; however, streptomycin and dihydrostreptomycin were detected at levels ranging from 695.85 to 1152.71 mg/kg and 6.35 to 11.11 mg/kg, respectively, in the culture media additives. The LC-MS/MS method coupled with direct addition of IPRs to the vial can provide useful basic data for AG analysis and safety evaluation of meats as well as culture media and additives for cultured meats.

• The Korean Journal of Pesticide Science
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• v.15 no.4
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• pp.545-572
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• 2011

The genetic diagnosis methods by RT-PCR and Virion capture (VC)/RT-PCR against Rice stripe virus (RSV) were developed. Three diagnosis methods of seedling test, ELISA and RT-PCR were compared in virus detection sensitivity (VDS) for RSV. The VDS of ELISA for RSV viruliferous small brown plant hopper (SBPH) was higher with 40.5% than that of seedling test. The VDS of RT-PCR was higher with 21% than that of ELISA. The VDS of ELISA and VC/RT-PCR was same with 9.2% in average on the SBPH collected from fields at the areas of Gimpo, Pyungtaeg and Sihueng, Gyeonggi province in 2009. The specific primers of RSV for SBPH and rice plant were developed for the diagnosis by Real time PCR. The RQ value of Real time PCR for the viruliferous and non viruliferous SBPH was 1 for 50 heads of non viruliferous SBPH, 96.5 for 50 heads of viruliferous SBPH, 23.1 for 10 heads of viruliferous SBPH + 40 heads of non viruliferous SBPH, and 75.6 for 30 heads of viruliferous SBPH + 20 heads of non viruliferous SBPH. The RQ value was increased positively by the ratio of viruliferous SBPH. Full sequences of 4 genomes of RSV RNA1, RNA2, RNA3 and RNA4 were analysed for the 13 RSV isolates from rice plants collected from different areas. Genetic relationships among the RSV isolates of Korea, Japan and China were classified as China + Korea, and China + Korea + Japan by phylogenetic analysis for RSV RNA1 and RNA2. In case of RNA3 involved in pathogenicity, genetic relationship of RSV among the three countries was grouped into 3 as China, China + Korea, and Korea + Japan. According to the genetic relationships in RSV RNA4, RSV isolates were grouped into 4 as China, Korea, China + Korea + Japan, and Korea + Japan. Viruliferous insect rate (VIR) of RSV in average increased in each year from 2008 to 2010, and the rates were 4.3%, 6.1%, and 7.2%, respectively, at the 28 major rice production areas in 7 provinces including Gyeonggido. The highest VIR in each year was 11.3% of Gyeonggido in 2008, 20.1% of Jellanamdo in 2009 and 14.2% of Chungcheongbukdo in 2010. The highest VIR depending upon the investigated areas was 22.1% at Buan of Jellabukdo in 2008, 36% at Wando and Jindo of Jellanamdo in 2009, and 30.0% at Boeun of Chungcheongbukdo in 2010. Average population density (APD) of overwintered SBPH was 13.1 heads in 2008, 13.9 heads in 2009 and 5.6 heads in 2010. The highest APD was 39.1 and 60.4 heads at Buan of Jellabukdo in 2008 and 2009, respectively, and 14.0 heads at Pyungtaeg of Gyeonggido. The acreage of RSV occurred fields was 869 ha in the western and southern parts, mainly at Jindo and Wando areas, of Jellanamdo in 2008. In 2009, RSV occurred in the acreage of 21,541 ha covered whole country, especially, partial and whole plant death were occurred with infection rate of 55.2% at 3,025 plots in 53 Li, 39 Eup/Myun, 19 Si/Gun of Gyeonggido, Incheonsi, Chungcheongnamdo, Jeollabukdo and Jeollanamdo. Seasonal development of overwintered SBPH was investigated at Buan, Jeollabukdo, and Jindo, Jeollanamdo for 3 years from 2008. Most SBPH developed to the 3rd and 4th instar on the periods of May 20 to June 10, and they developed to the adult stage for the 1st generation on Mid and Late June. In 2009, all SBPH trapped by sky net trap were adult on May 31 to June 1 at Mid-western aeas of Taean, Seosan and Buan, and South-western areas of Sinan and Jindo. The population density of adult SBPH was 963 heads at Taean, 919 at Seocheon and 819 at Sinan area. The origin of these higher population of adult SBPH were verified from the population of non-overwintered SBPH but immigrant SBPH. From Mid May to Mid June in 2010, adult SBPH could not be counted as immigrant insects by sky net trap. The variation of RSV VIR was high with 2.1% to 9.5% for immigrant adult SBPH trapped by sky net trap at Hongsung of Chungcheongbukdo, Buan of Jeollabukdo and so forth in 2009. The highest VIR for the immigrant adult SBPH was 9.5% at Boryung of Chungcheongnamdo, followed by 7.9% at Hongsung of Chungcheongnamdo, 6.5% at Younggwang of Jeollanamdo, and 6.4% at Taean of Cheongcheongnamdo. The infection rate of RSV on rice plants induced by the immigrant adult SBPH cultivated near sky net trap after about 10 days from immigration on June 12 in 2009 was 84.6% at Taean, 65.4% at Buan and 92.9% at Jindo, and 81% in average through genetic diagnosis of RT-PCR. Barley known as a overwintering host plant of RSV had very low infection rate of 0.2% from 530 specimens collected at 10 areas covering whole country including Pyungtaeg of Gyeonggido. Twenty nine plant species were newly recorded as natural hosts of RSV. In winter annual plant species, 11 plants including Vulpia myuros showed RSV infection rate of 24.9%. The plant species in summer annual ecotype were 13 including Digitaria ciliaris with 44.9%, Echinochloa crusgalli var. echinata with 95.2% and Setaria faberi with 65.5% in infection rate of RSV. Five perennial plants including Miscanths sacchariflorus with infection rate of 33.3% were recorded as hosts of RSV. Rice cultivars, 8 susceptible cultivars including Donggin1 and 17 resistant ones including Samgwang, were screened in field conditions at 3 different areas of Buan, Iksan and Ginje in 2009. All the susceptible cultivars were showed typical symptom of mosaic and wilt. In 17 genetic resistant cultivar, 12 cultivars were susceptible, however, 5 cultivars were field-resistant plus genetic resistant to RSV as non symptom expression. When RSV was artificially inoculated at seedling stage to 4 cultivars known as genetic resistant and 3 cultivars known as genetic susceptible, the symptom expression in resistant cultivars was lower as 19.3% in average than that of 53.3% in susceptible ones. In comparison of symptom expression rate and viral infection rate using resistant Nampyung and susceptible Heugnam cultivars by artificial inoculation of RSV at seedling stage, the symptom expression of Heugnam was higher as 28% than 12% of Nampyung. However, virion infection of resistant Nampyung cultivar was higher as 12% reversely than 85% of susceptible Heugnam. Yield loss of rice was investigated by the artificial inoculation of RSV at the seedling stage of resistant cultivars of Nampyung and Onnuri, and susceptible cultivars of Donggin1 and Ungwang for 3 years from 2008. The average yield per plant was 7.8 g, 8.5 g and 13.8 g on rice plants inoculated at seedling stage, tillering stage and maximum tillering stage, respectively. The yield loss rate was increased by earlier infection of RSV with 51% at seedling stage, 46% at tillering stage and 13% at maximum tillering stage. In resistant rice cultivars, there was no statistically significant relation between infection time and yield loss. In natural fields on susceptible rice cultivar of Ungwang at Taean and Jindo areas in 2009, the yield loss rate was increased with same tendency to the infection hill rate having the corelation coefficient of 0.94 when the viral infection was over 23.4%.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.4 no.1
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• pp.115-124
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• 1968

It was shown that the most desirable characters for kenaf are high-fiber weight and moderately early maturity. Therefore, the objectives of this research on this crop is to find varieties possessing these characteristics. The experiments covered in this report provided new information relative to segregation, mode of inheritance, estimate of the number of genes involved in fiber weight and their response to short day length of 10 hours and the qualitative characters, such as, color of stem, capsule, petiole and shape of leaves. The associations which exist among these characters are also indicated. Fiber weight per plant, days to flowering, Stem color, Petiole color, Capsule color, and shape of leaves were studied in parental, $F_1$.$F_2$and backcross populations of a cross between Dashkent, a low-fiber weight but early maturing kenaf variety, and G 38 F-1, a high-fiber weight but late maturing kenaf variety. Crosses were made using the varieties, Dashkent and G 38 F-1 as parents. The Dashkent parent had the following characteristics: green stems, capsules and petioles and lobed shaped leaves; 105.8234 mean-days to flowering in the field, and 106.9222 mean-days under 10 hours short day treatment. The other parent, G 38 F-1 had red stems yellow capsules and red petioles and unlobed shaped leaves; 149.8921 mean-days to flowering in the field, and 62.3684 mean-days under 10 hours short day treatment. Both of the parents, $F_1$, $F_2$, $BC_1$ ($F_1$ X Dashkent, ) and $BC_2$($F_1$ ${\times}$ G38F-1) of the kenaf cross were grown at the Crops Experiment Station, Suwon, Korea in 1965. Color of stems, petioles and capsules, and shape of leaves were noted to be simply inherited as a single factor. Red stem color was dominant over green stem color, red petiole color was dominant over green petiole, lobed shaped leaves were dominant over unlobed shaped leaves and yellow capsules were dominant over green capsule. It was, also, noted that the factor for color of petiole was linked with the factor for shape of leaf with a 11.9587 percent recombination value, however no interaction or linkage were found among the color of stem and capsule color. Using Powers partitioning method, theoretical means and frequency distributions for each population, the days to flowering were calculated with the assumption that two gene pairs were involved. The values obtained fitted the theoretical values. In general this would indicate that Dashkent and G 38 F -1 were differentiated by two gene pairs. Heritability values were calculated as the percent of additive genetic variance. Heritability value of days to flowering, 89.5% in the broad sense and 79.91% in the narrow sense, indicated that the selection for this character would be effective in relatively early generations. Particularly, high positive correlations were found between days to flowering and the color of petioles and shape of leaves. However, there was no relation between days to flowering and capsule color nor between these and stem color. On the basis of the results of this experiment there is evidence that the hereditary factor for shape of leaves and the color of petioles is linked with an effective factor or factors for the characters of days to flowering. The association was sufficiently close to offer a possible simple and efficient means of selection for moderately early mat. uring plants by leaf shape and petiole color selection. Again using Powers partitioning method the frequency distribution for each population to the fiber weight were calculated with the assumption that two gene pairs, AaBb, were involved. Both phenotypic and genotypic dominance were complete. The obtained value did not agree with the theoretical value for $F_2$ and $BC_1$ ($F_1$ ${\times}$ Dashkent.) It seems that Dashkent and G 38 F-1 were differentiated by two major gene pairs but some the other minor genes are necessary. It is certain that the hereditary factor for shape of leaves and color of petioles is linked with an effective factor or factors for fiber weight. Also, high. yielding plants with moderately early maturity were found in the $F_2$ population. Thus, simultaneous selection for high-fiber yield and moderately early maturing plants should be possible in these populations. Phenotypic and genotypic correlation coefficients between fiber weight per plant and days to flowering, stem height and stem diameter were calculated. In general, genotypic correlations are higher than the phenotypic correlation. The highest correlation is found between stem height and fiber weight per plant (0.7852 in genotypic and 0.4103 in phenotypic) and between days to flowering and fiber weight per plant (0.7398 in genotypic and 0.3983 in phenotypic.) It was also expected that the selection of high stem height and moderately early maturing plants were given the efficient means of selection for high fiber weight.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.4 no.1
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• pp.31-71
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• 1968

To clarify the breeding behavior of the hybrids between tropical and temperate area rice varieties, investigations were made on heading days and grain sterility. In this study, crosses were made in half way diallel involving 7 varieties: 2 photoperied sensitive Indicas, 2 less sensitive intermediate Indicas, 1 Ponlai Japonica and 2 high temperature sensitive Japonicas. The parents and $F_1$s were grown under 10 hours and 14 hours daylength controlled conditions at both IRRI(International Rice Research Institute, N$14^{\circ}$17') and Suwon(N$37^{\circ}$16'). F2s with their parents were grown at IRRI in the short day season, and at Suwon under natural conditions. Fa lines with their parents were grown at Suwon under natural conditions. Observations were made for heading days and sterility. The results are summarized as follow; 1. Heading days : 1. For the $F_1$s, earliness showed dominance or overdominance to lateness under the 10 hours condition, and dominance or partial dominance under the 14 hours conditions, at both IRRI and Suwon. 2. For the $F_2$s grown at IRRI during the shortday season earliness appeared to be dominant over lateness and segregation was not distinct and continuous. In the early season culture of $F_2$s at Suwon earliness showed partial dominance or was intermediate. In the proper season culture of $F_2$s lateness showed partial dominance or was intermediate. 3. In the combinations between late parental varieties which do not head at Suwon, transgressive segregants bearing effective panicles were obtained. 4. The crosses of parental varieties having long basic vegetative growth duration showed bigger variance in heading days, and significant correlation was found between of parental varieties and the mean coefficient of variance for parental arrays. 5. The means of heading days of F2 populations were significantly correlated with those of $F_1$ or mid-parents. The means of F 8 lines were also highly correlated with the means of $F_2$s, but, the means of $F_3$ lines grown at Suwon and of their parental $F_2$ individual, grown at IRRI were not correlated. 6. A faint heritability was calculated from the regression of $F_3$ lines grown at Suwon on the $F_2$ individuals grown at IRRI for most combinations, especially in the combinations involving shortday sensitive varieties. This implies low efficiency for the selection of heading days of $F_2$ individuals at IRRI to be grown in lines at Suwon. 7. No significant reciprocal effects were measured for $F_1$ and $F_2$ mean heading days. 8. Partitioning the observed photoperiod sensitivity. into two components, parental array mean md the deviation from this array mean, the parental photoperiod sensitivity contributing to the hybrids was measured in terms of general and specific combining ability for photoperiod sensitivity. 9. The photoperiod sensitivity of $F_1$s was higher than that of the parents, and it decreased as the generation progressed in most combinations of tested varieties. 10. The response of heading days to difference of temperature was weaker for $F_1$ hybrids than for the parents. The differences of temperature responses between the longday and shortday treatments were specific for the variety. 2. Sterility : 1. The $F_1$ sterility was specific for the combinations and not correlated to the parental sterility. The sterility of $F_1$s grown under the 10 hours condition was higher than of those grown under 14 hours. These results were the same at both locations, IRRI and Suwon. 2. The high sterile combinations in $F_1$ showed high sterility in $F_2$. The combinations between a high photoperiod sensitive variety and a high temperature sensitive variety showed high sterility and wider variance. 3. The mean sterility of $F_2$s was lower than of $F_1$s and the mean of $F_3$ lines was lower than of $F_2$s. Sterility decreased as the generation progressed, and the differences of $F_3$ sterility of different combinations were not significant. 4. A faint correlation between grain sterility and pollen sterility was observed in $F_2$ populations. 5. No significant reciprocal effects were measured in $F_1$ and $F_2$ sterility. 6. Following Griffing's method, specific combining ability effects were higher than general combining ability effects, especially in the combinations between highly photoperiod sensitive varieties and highly temperature sensitive varieties. 7. No distinct correlations were found between $F_2$ individual sterility grown at IRRI and $F_3$ line sterility grown at Suwon. 8. No distinct correlations were observed between heading days and sterility of $F_2$ individuals.

• Journal of Korean Society of Forest Science
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• v.16 no.1
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• pp.1-32
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• 1972

By means of the interspecific hybridization in the Sub-genus Diploxylon of the Genus Pinus, $F_1$ hybrids of Pinus rigida${\times}$elliottii, Pinus rigida${\times}$radiata, P. rigida${\times}$serotina and P. densiflora${\times}$thunbergii had been produced. And on the basis of the crossabilities of these hybrids the taxonomic affinities of these pines were examined. And the needle characteristics of these hybrid and the occurence of phenolic substances in these $F_1$ hybrid were also investigated to see the potential usefulness of these characteristics for the diagnosis of the taxonomic affinity. And, the growth performances of the $F_1$ hybrids have also been compared with those of parental species. In order to contribute to the establishment of the hybrid seed orchard the introgression phenomena between P. densiflora and P. thunbergii in the eastern coastal area have also been investigated along with the investigation of the heterozygosity of plus trees of P. densiflora growing in the clone bank in Suwon. And the results were summarized as follows. 1. On the basis of crossabilities as well as on the taxonomic affinities according to the systems of Shaw, Pilger and Duffield, it has been proven that the parental species of those hybrids are of close affinities and range of the fertile hybrid seed production rate was as high as 28-58% in the best hybrid combination (Table 13). 2. Among those hybrids, the ${\times}$ Pinus, rigiserotina hybrid seemed to be most promising in the growth performance exhibiting 109-155% more volume growth compared to the seed parent with the statistic significance of 1% level (Tables 16 and 17). 3. Notwithstanding the fact that the all of the pollen parents are cold tender, all hybrids exhibit cold hardiness as much as their seed parent and it seems to suggest that the characteristics of cold hardiness were transmitted from the seed parent. 4. Though a striking difference in needle length was observed between the parental species of each hybrid, it was difficult to distinguish each hybrid from their seed parent by the needle length except ${\times}$P. rigiserotina which is characterized by long needle which is 65% more longer than the needle of the seed parent (Table 21). 5. With regard to the anatomical characteristics of needle, the hypoderm is apparently thicker in most of the $F_1$ hybrid pines and the characteristics of resin canals are dominated by medial in most $F_1$ hybrid. And, the fibrovascular bundles were apart as were in their seed parent. Therefore it was found to be possible to distinguish the hybrids pines from their parents by the needle characteristics. And, it is to be noticed that the ${\times}$P. densithunbergii was more close to the pollen parent having RDI value of 0.73 (Fig.l, Table 22). 6. It has been demonstrated that ${\times}$P. rigielliottii, ${\times}$P. rigiradiata and ${\times}$P. rigitaeda have a phenolic substance (No.7) of light yellow at Rf-0.46, same as their seed parent, but no trace of phenolic substance was observed in their pollen parent. This fact will serve as an important criteria for early identification of hybridity in progeny testing. However, the fact that both of ${\times}$P. rigiserotina and ${\times}$P. densithunbergii exhibit the same reactions of phenolic substances as well their parental species seems to indicate the close affinities between the parental species of the respective hybrid (Fig.2, Table 23). 7. The separation and the reaction of phenolic substance developed on TLC were found to be same in the same species showing no variations between the individuals, and no variations due to tree part of sampling, tree age or pollen sources. And the reaction was also observed regardless of the not varied by the kind of developing solvent whether it is Aceton-Chloroform (3:7 v/v) or Benzene-Methanol-Acetic acid (90:16:8 v/v). 8. The introgression phenomena of natural Pinus densifiora stand in both east and west coastal area indicates that the major part of the red pines investigated are all heterozygous and the heterozygosity of pines are higher in the west coast than in the east coast(Tables 24 and 25). 9. Based on the RDI, among the plus trees of Pinus densiflora selected in Korea and Japan as well, no pure P. densiflora has been found. Since all of the sample trees of Pinus densiflora were found to be as heterozygous bearing part of the characteristics of P. thunbergii, those red pines were considered to be natural heterotic hybrid pines(Figs. 3 and 4. Tables 26 and 27).