Qualitative and quantitative analysis of flavonoids from 12 species of Korean mulberry leaves.

The total flavonoids in leaves of 12 varieties of Korean mulberry (Morus alba L.) were determined. Seventeen flavonoids were isolated and analyzed using ultra-performance liquid chromatography coupled with diode array detection and quadrupole time-of-flight mass spectrometry (UPLC-DAD-QTOF/MS). To determine the flavonoid contents, HPLC analysis was performed on these 17 flavonoids. The total flavonoid contents of the 12 varieties of mulberry leaves ranged from 748.5 to 1297.9 mg, with the highest obtained from the Cheong Su variety (1297.9 ± 112.0 mg). Among the 17 flavonoids analyzed, quercetin 3-O-rutinoside (rutin) and quercetin 3-O-glucoside (isoquercitrin) had highest contents in the Cheong Su variety. Furthermore, the Dae Dang Sang variety gave the highest quercetin 3-O-rutinoside (rutin) content among the mulberry leaves investigated, at 425.5 ± 45.9 mg. Major flavonols from Dae Dang Sang were detected by UPLC-DAD-QTOF/MS. A total of 17 flavonoid compound peaks were identified in the analysis time range of 5-40 min, all of which were kaempferol and quercetin glycosides. Seven of the 17 compounds identified in mulberry leaves were unknown.

Accumulation of Rutin and Betulinic Acid and Expression of Phenylpropanoid and Triterpenoid Biosynthetic Genes in Mulberry (Morus alba L.).

Mulberry (Morus alba L.) is used in traditional Chinese medicine and is the sole food source of the silkworm. Here, 21 cDNAs encoding phenylpropanoid biosynthetic genes and 21 cDNAs encoding triterpene biosynthetic genes were isolated from mulberry. The expression levels of genes involved in these biosynthetic pathways and the accumulation of rutin, betulin, and betulinic acid, important secondary metabolites, were investigated in different plant organs. Most phenylpropanoid and triterpene biosynthetic genes were highly expressed in leaves and/or fruit, and most genes were downregulated during fruit ripening. The accumulation of rutin was more than fivefold higher in leaves than in other organs, and higher levels of betulin and betulinic acid were found in roots and leaves than in fruit. By comparing the contents of these compounds with gene expression levels, we speculate that MaUGT78D1 and MaLUS play important regulatory roles in the rutin and betulin biosynthetic pathways.

Synnemata Production Using Silkworm Variety, Female Yangwonjam by Isaria tenuipes.

This study was conducted to confirm the possible use of female Yangwonjam as a host for synnemata production of Isaria tenuipes in eight local areas in Korea. Silkworm pupation rate, infection rate and synnemata characteristics of I. tenuipes were examined. Normal silkworms had a higher pupation rate than silkworms inoculated with I. tenuipes. The pupae survival percentage of normal silkworm in cocoons was 92.5~97.6%, whereas it ranged from 91.1~95.6% in silkworms sprayed with I. tenuipes. Female Yangwonjam showed the highest survival percentage at 97.6% among the silkworm varieties tested. I. tenuipes infection rate of larvae of 5th instar newly-exuviated silkworms was 89.2~90.7% in the spring rearing season and 98.2~99.3% in the autumn rearing season. Synnemata production of I. tenuipes was excellent in female Yangwonjam with an incidence rate of 98.0% followed by male Yangwonjam (94.1%) and Baegokjam (93.3%) in the spring rearing season. Synnemata living weight ranged from 1.44~0.94 g in the spring rearing season. The female Yangwonjam had the heaviest synnemata weight (1.44 g) in the spring rearing season. The synnemata of I. tenuipes produced on pupae were white or milky-white in color, and were similar in shape and color to wild synnemata collected in Korea.

Breeding of a Silkworm Variety for Synnemata Production of Isaria tenuipes.

This study was conducted out to select a silkworm variety suitable for synnemata production of Isaria tenuipes. Four kinds of the mulberry silkworm varieties, Bombyx mori, were hybridized using a Japanese parental line and a Chinese parental line, and used to test for synemata formation in I. tenuipes. The larval period of normal silkworms was 22 hr longer than the silkworms inoculated with this fungus. Among the silkworm varieties tested, Hachojam had the shortest larval period with 23.02 days. The non-cocooning silkworm had a shorter larval period than the cocoon producing silkworms. The pupation rate of normal silkworms was about 9% higher than that of silkworms sprayed with I. tenuipes. Hachojam had the highest infection rate at 99.8%, but no significant difference was observed for the infection rate by silkworm variety. The production of synnemata was the best in JS171 × CS188 with an incidence rate of 99.3%, followed by Hachojam, and Chugangjam. The synnemata produced from Hachojam were the heaviest and showed white or milky-white in color.

Identification and distribution of two fungal species causing sclerotial disease on mulberry fruits in Korea.

A total of 520 overwintered sclerotia were collected from surface of soil under mulberry trees in six locations in Korea during February in 2006 and 2007. The collected sclerotia were tested for their germination in vitro and identified based on their morphological characteristics. Out of all sclerotia tested, 52.3% of the sclerotia germinated and produced two types of apothecia. The two types of fungi occurred from the sclerotia at the ratio of 49.8 vs. 50.2. The fungal type with cup-shaped apothecia was identified as Ciboria shiraiana and another type of fungus with club-shaped apothecia as Scleromitrula shiraiana. Taxonomy and distribution of the two sclerotial fungi were described and discussed.

Chemical Components of Paecilomyces tenuipes (Peck) Samson.

The caterpillar-shaped Chinese medicinal mushroom (DongChongXiaCao) looks like a worm in the winter and like a grass in the summer. The fruiting body has been regarded as popular folk or effective medicines used to treat human diseases such as asthma, bronchial and lung inflammation, and kidney disease. The fruiting bodies of Paecilomyces tenuipes that formed on the living silkworm (Bombyx mori) host were used in this examination. This study was carried out to investigate the proximate composition, soluble sugar, amino acid and fatty acid profiles, and contents of the bioactive ingredient including adenosine and D-mannitol in the fruiting-bodies. The moisture content was 57.56%. Soluble sugars found were glycerol, glucose, mannitol and sucrose, and the contents exceeded 24 mgg(-1)dry weight. Total free amino acid content was 17.09 mg g(-1)dry weight. Arginine, glycine, proline and tyrosine were main amino acids. The content of oleic acid in fatty acids was high. Adenosine was more abundant in fruiting bodies than corpus.