• The Korean Journal of Soil Zoology
• /
• v.8 no.1_2
• /
• pp.23-26
• /
• 2003

A neem based product, Suneem-G (granules) alone as well as in combination with urea was tested against root-knot nematode, Meloidogyne incognita, on okra (Abelmoschus esculentum) in clay pots of 20-cm-diameter, containing 2 kg sterilized soil, inoculated with 2,000 freshly hatched second stage juveniles (J$_2$) of M. incognita. Application of different dosages of Suneem-G with or without urea significantly reduced nematode population and incidence of galling on okra roots and increased vegetative plant growth compared with the untreated control. Suneem-G with urea was the most effective and its effect increased with the increase of dosage. Suneem-G 6 g＋urea 25 g per pot gave maximum plant growth and root gall reduction, as plants shoot and root lengths were increased by 84 and 58% and root galling was reduced by 58%, respectively. The lowest dosage of Suneem-G alone, 2 g per pot increased shoot and root lengths of plants by 25.7 and 17%, respectively, and reduced root galling by 24%. The lowest effect was recorded in the pots treated with 25 g urea alone; shoot and root lengths were increased and root galling was decreased by 14% each.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.1
• /
• pp.23-28
• /
• 1994

This study was rimed out to examine the difference in the cell vitality between mesophyII protoplast (MP) and paraveinal mesophyII protoplast (PVMP) of Nicotiana tabaccum 'Xanti', Petunia hybrida 'Blue Star' and Chrysanthemum morifolium 'Baeckwang' by using urea permeability technique. The effects of various enzyme solutions and incubation time, NAA and thidiazron on plant regeneration from isolated protoplasts were also investigated. The vibratome technique was used for protoplast isolation and urea permeability test because the fresh living, thin tissue stripes (50 ${\mu}{\textrm}{m}$ of thickness) could be obtained with minimal damage with the vibratome. For the three plants examined, the urea permeability on the tested tissue stripes was relatively higher in PVMP than in MP by about Ks = 2.0 $\times$ 10$^{-5}$ cm/sec. The treatment of an enzyme mixture of 1.5% cellulase R-10, 1% Driselase, 0.5% Macerozyme R-10, and 0.5% Pectinase for 4 to 8 h was effective on the isolation of PVMP. The highest frequency of callus formation and plant regeneration from the isolated protoplasts was obtained with NAA 2 mg/L and thidiazuron 0.01 mg/L. Furthermore, the results demonstrated that cell devision and plantlet regeneration was more frequent in the PVMP than in the MP of the same leaf or plant We, therefore, conclude that UM is an excellent experimental material for the callus formation and regeneration from isolated protoplasts.

• Applied Microscopy
• /
• v.40 no.2
• /
• pp.101-108
• /
• 2010

Vitex negundo is an aromatic plant which releases a unique scent due to the presence of essential oil stored presumably within glandular trichomes. The focus of this research was to study developmental patterns of glandular trichomes in Vitex negundo leaves using electron microscopy. There are two types of glandular trichomes which develop on the leaf epidermis of Vitex negundo, peltate glandular type (PT) and capitate glandular type (CT). Structural features differ significantly depending on size and density, formation of secretory cavity, plastid, etc during developmental stages. In young leaves, undifferentiated PTs are densely distributed in the upper epidermis, but are not externally exposed in the lower epidermis because they are covered by non-glandular simple trichomes. Upon leaf development, PTs and CTs show clear structural differentiation in the upper and lower epidermis. PTs are composed of up to eight head cells (ca. 35~40 ${\mu}m$) and one stalk cell (ca. 5 ${\mu}m$), while CTs are composed of four head cells (ca. 10~15 ${\mu}m$) and 1~2 stalk cells (ca. 10 ${\mu}m$). Although secretory cavities develop on the secretory head cells, their size, structure, and formation proceed very differently depending on trichome type. In early development of PT, a large cavity with numerous secretory vesicles form rapidly from the head cells. In CT, however, only a small secretory cavity is formed, slowly relative to PT, without secretory vesicles. The PTs are considered to play an important role in releasing the aromatic components of Vitex negundo.

• Korean Journal of Agricultural Science
• /
• v.13 no.2
• /
• pp.157-167
• /
• 1986

The study was carried out to identify several factors affecting isolation and culture of cotyledone and leaf mesophyll protoplasts of cabbage (Brassica oleraceae), petsai (B.campestris subsp. pekinensis) and rape (B.napus). High viable protoplasts could be obtained when the cotyledon and the leaf mesophyll tissue of all species were treated with enzyme solution composed of 1% macerozyme 'R-10', 1.5% Onozuka 'R-10', 10% mannitol and 50,0 mM $CaCl_22H_2O$ for 4 hours. The protoplasts which obtained from the cotyledon of all species except the cabbage and the leaf mesophyll tissue of all species were divided on NN culture medium supplemented with 9.1% mannitol, 1% glucose, 1% sucrose, $1mg/{\ell}$ 2,4-D, $0.5mg/{\ell}$ NAA and $0.5mg/{\ell}$ BA. The division of the rape leaf mesophyll protoplasts were continued and led to colony.

• Korean Journal of Agricultural Science
• /
• v.12 no.1
• /
• pp.22-30
• /
• 1985

Protoplasts of Pisum sativum L. were isolated and cultured from leaf mesophyll tissue. The successful yield of protoplast was obtained in an enzyme solution of 2% 'Onozuka R-10' and 2 % 'Macerozyme R-10' contained 6mM $CaCl_2{\cdot}2H_2O$ within 4 hours. They were divided in B5 culture medium supplemented with 2mg/l kinetin, 1mg/l 2, 4-D and 0.2% Difcobacto agar. Divisions of the protoplasts were continued and led to colony formation for 1 months. The colony from protoplasts of pea mesophyll tissue was formed to callus after subculture in a medium contained macronutrients and amino acids of BII medium and micronutrients and vitamins of B5 medium, and also supplemented with 2mg/l kinetin 2mg/l NAA.

• Korean Journal of Plant Taxonomy
• /
• v.44 no.2
• /
• pp.119-131
• /
• 2014

A comparative study of leaf anatomy in tribe Sorbarieae (Adenostoma, Chamaebatiaria, Sorbaria, Spiraeanthus) including one related genus Lyonothamnus was carried out using light microscopy. Anatomical characteristics of the leaf blade and midrib were described and taxonomically evaluated. The anatomical characters which described in this study are as follows: thickness of leaf midrib, blade in cross section, cuticle, epidermal cell, stoma, trichome, mesophyll, crystal, main vasculature type. All features were compared and the vascular patterns of midrib were distinguished two types. - Type 1: Trace tripartite (Adenostoma), Type 2: Trace continuous, subtype 2A: flat arc (Chamaebatiaria, Spiraeanthus), subtype 2B: U-shape arc (Lyonothamnus, Sorbaria). In conclusion, some of leaf anatomical characters (e.g., cuticle, epidermal cell, trichome, mesophyll, main vasculature type) can be useful for diagnostic features. Hypostomatic type, dorsiventral mesophyll, Ushape vasculature type would constitute a major characters for genus Sorbaria in Sorbarieae. The detailed anatomical description of studied taxa is provided, and its systematic importance is also briefly discussed.

• Journal of Yeungnam Medical Science
• /
• v.2 no.1
• /
• pp.175-181
• /
• 1985

Agrobacterium tumefaciens induces cancerous growths called crown galls at wound sites on dicotyledonous plants. A large plasmid called Ti plasmid is responsible for virulence. Upon tumor induction, part of the plasmid, termed T-DNA, becomes integrated into plant genome and its genetic sequences are expressed. These properties allow Ti plasmids to be used as gene vectors in plants. Several in vitro methods for the transfer of Ti plasmid into plant cell have been developed. One of them is the treatment of bacterial spheroplasts and plant protoplasts mixture with polyethylene glycol that is generally used as fusogen in cell-to-cell fusion. Several workers investigated the interaction of bacterial spheroplasts with plant protoplasts in the presence of polyethylene glycol and suggested that the interaction is not fusion but endocytosis. In this report we observed the interaction of Agrobacterium tumefaciens spheroplasts with Nicotiana tabacum protoplasts by electron microscope. Agrobacterium tumefaciens spheroplasts and Nicotiana tabacum protoplasts were prepared and mixed in the presence of polyethylene glycol and high pH-high $Ca^{2+}$ buffer. Then the interaction of the spheroplasts with the protoplasts was examined by transmission electron microscope. After the treatment of polyethylene glycol the spheroplasts adhered to the surface of the protoplasts and then they were engulfed by the protoplasts. After the high pH-high $Ca^{2+}$ buffer treatment the engulfed spheroplasts lost their cell integrity. No fusion process was observed. Thus all these observations suggest that the introduction process of Agrobacterium tumefaciens spheroplasts into Nicotiana tabacum protoplasts with the aid of polyethylene glycol is endocytosis.

• Korean Journal of Weed Science
• /
• v.16 no.3
• /
• pp.237-244
• /
• 1996

Propanil [N-(3,4-dichlorophenyl) propanamide] which was applied at 4,200g ai/hapostemergence 7 days after seeding or transplanting, completely reduced the growth of shoot and root of barnyardgrass at 100% under dry condition while plant height, root length and shoot fresh weight of barnyardgrass at 63, 40 and 78%, respectively under water condition. On the other hand, the herbicide did not affect the growth of shoot and root of rice grown under water condition and transplanting condition, but reduced the plant height, root length and shoot fresh weight of broadcast rice on soil at 24, 18 and 28%, respectively, under dry condition. Microscopically, the epidermal cells of treated-barnyardgrasses under both conditions were severely constricted, chloroplasts in the cells of vascular bunble sheath were partially lacked, and mesophyll cells were often ruptured, whereas those of treated-rice were not affected. Histological observations showed that propanil reduced the thickness of leaf blade of barnyardgrass under both conditions at 36-48% due to mainly reduction and constriction of mesophyll cell, while it did not affect or even increased the thickness of leaves of rice under all conditions compared to control. These results indicate that broadcast rice on soil were more injured than drilled rice in soil under dry condition, however, in the other tested conditions ricer were not affected.

• Current Research on Agriculture and Life Sciences
• /
• v.2
• /
• pp.15-22
• /
• 1984

This study was conducted to identify the enzyme treatment time, enzyme concentration and plant growth condition for isolation of potato mesopyll, it was also performed to determine the adquate sucrose molarity on purification of protoplasts and to investigate the incubation time, PEG concentration and DMSO effect for potato-petunia protoplast fusion. The results were summarized as follows: The optimal time of incubation in enzyme solution was 3 - 4 hours and high humidity and low light intensity made plants more effective to protoplast releasing enzymes. Our experimental results showed that the pectolyase Y-23 was an ideal agent for isolation from mesophyll cultured in vitro compared with macerozyme. The enzyme solution with 0.5 % macerozyme and 2 % cellulase was very effective and the purity of healthy protoplast was better in 0.4 and 0.5 M sucrose than in others. It was revealed that the rate of potato-petunia fusion according to the incubation time with PEG was effective at 30 min incubation and percentage of protoplast aggregation was increased by high molecular weight and concentration of PEG. Percentage of potato-petunia protoplast heteroplasmic aggregation was increased by 4 to 16 % in PEG 6000 compared with PEG 4000 and PEG 1500. Addition of 5 to 10 % DMSO to the PEG solution increased to the heteroplasmic aggregation of potato-petunia from 2 to 4 %.

• Journal of Korean Society of Forest Science
• /
• v.73 no.1
• /
• pp.33-42
• /
• 1986

This study was carried out to investigate the optimum conditions for isolation and culture of mesophyll protoplasts from Populus alba ${\times}$ P. glandulosa. The results obtained from the experiments are as follows; 1) The suitable concentration of BAP for shoot multiplication was 0.4 mg/l. 2) High yield and viability of isolated protoplasts were obtained by our high enzyme-short time incubation method. 3) Optimum enzyme concentrations for mesophyll protoplast isolation were Cellulase 2%, Macerozyme 0.8%, Hemicellulase 1.2%, Driselase 2%, and Pectolyase Y-23 0.05%. 4) 0.6M mannitol in enzyme solution was the most effective for protoplast isolation and viability. 5) The most adequate pH level of enzyme solution was pH 5.6. 6) The effect of DTT and MES buffer was significant. 7) For protoplast purification, 0.6M sucrose was the most proper concentration. 8) The adding effect of Dextran T40 in floating solution was important. 9) The mesophyll protoplasts isolated through our high enzyme-short time incubation method revealed successful response to culture condition over 3 weeks of culture.