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Sulaymonov Botirjon Abdushukirovich, Academician, Rector of Tashkent State Agrarian University, Kimsanbaev Xojimurod Xamraqulovich, Professor, Department of Plant protection from pests and quarantine,
Jumaev Rasul Axmatovich, Ph D., Department of Plant protection from pests and quarantine,
Sabirov Sobir Karimbaevich, Master degree, Department of Plant protection from pests and quarantine, E-mail: [email protected]; [email protected]
REARING OF TRICHOGRAMMA SPECIES (T. EVANESCENS, T. PINTOI, T. CHILONIS) IN VITRO CULTURE
Abstract: This article reports Trichogramma evanescens, Trichogrammapintoi and Trichogramma chilonis (Hymenoptera: Trichogrammatidae) were cultured in vitro from eggs to adults on artificial diets, which contained no insect components. The results of our experiments using artificial media devoid of insectan additives for rearing T. evanescens, T.pintoe, T.chilonis in vitro.
Keywords: Artificial media, parasitoid growth factors, in vitro, hemolymph, egg yolk, inorganic salt mixture, cow milk, Trichogramma spp, predator, biological efficiency.
Introduction Okkurgan and Piskent districts, and reared in the labora-
At the process of entirely rearing trichogramma para- tory on Heliothis armigera and Agretus segetum. site in the laboratory condition cultivates Sitotrogo cere- Ingredients for in vitro medium: Pupae hemo-
allela Olive egg a lot of corn productions such as grain, maize, barley and worker force are spent. In the result of it, rearing price is increased. More than 150-thousand-ton barley is emitted in exist biolaboratories in Uzbekistan in a year [4; 5; 6].
Emitting this barley, other diets cultivation happens in the account of decreasing producing of plants or enterprises. This testifies that above mentioned process is damaging for diets industry.
In Uzbekistan 2014-2016 years for 3 year' investigations rearing trichogramma in the diets medium, which belongs to artificial medium researches showed its good results. (Jumaev R., 2016., 2017).
The artificial rearing of Trichogrammatidae started a long time ago, with the main goal to try obtaining a mean to multiply and produce parasitoids to be released in biological control strategies. But it is also a powerful tool to conduct studies on biology, physiology and behavior of entomophages, especially endoparasitoid species [1; 3; 5].
Materials and methods
Trichogramma stock: T. evenecens, T.pintoe, T.chilonis were collected from Tashkent province, Buka,
lymphy (G.melonnellan or H.armigera), cow milk or 10% powdered milk solution, chicken embryo extract and Neisenheimer's mixture salt.
Preparation of the ingredients
- Insect hemolymph collection: A live pupa was immersed in water bath at 60 ° C for 6 or 7 min to avoid blackening of the hemolymphy. After surface sterilization with alcohol and need sterile condition.
Chicken embryo extract collection: The Chicken embryo extract, only it should be the egg yolk and need sterile condition (Jumaev R., 2016., 2017).
- Milk: Fresh cow milk or 10% powdered milk solution also need sterile condition.
- Inorganic mixture salt: Use Neisenheimer's mixture salt (NaCl 7.5 g, KCl 0.1 g, CaCl2 0.2 G, NaHCO3 0.2 g, H2O 100 ml).
Artificial "egg-cards"
There are 2 types of artificial "egg-cards Tri-ring "eggcards" 2 pieces of plastic film are used. The semispherical concaves are made on the upper plastic film. Artificial medium is poured into concaves fully (but without overflow) with a micro-syringe or micro-pipette. The bottom
Section 1. Biology
plastic film has no concaves. The upper and bottom plastic films are separated and stretched tightly by three plastic ring with different inner diameters, in our Cass they are: 5:5 cm, 5.4 cm, 5.2 cm respectively [1; 2; 4].
Components of artificial diets for 3 Trichogram-ma spp.
The first medium: for Trichogramma evanescens, He-molymph (A) G.melonnellan (A1) 41.5 ± 0.5%, Neisen-heimer's (A2) 15.5 ± 0.5%, Chicken egg yolk (A3) 20.5 ± ± 0.5%, milk (A4) 22.5 ± 0.5%.
The second medium: for Trichogramma pintoi, He-molymph (B) Heliothis armigera (B1) 45.5 ± 0.5%, Neis-enheimer's (B2) 13.5 ± 0.5%, Chicken egg yolk (B3) 20.5 ± ± 0.5%, milk (B4) 20.5 ± 0.5%.
Table 1. - Development of Trichogramma reared in vitro laboratory experiences
The three medium: for Trichogramma chilonis, He-molymph (C) Agretus segetum (C1) 45.2 ± 0.5%, Neisen-heimer's (C2) 13.2 ± 0.5%, Chicken egg yolk (C3) 21.6 ± ± 0.5%, mill2 (C4) 20.0 ± 0.5% [2; 4; 5; 6]. The results of the research It was showed in the Table 1 that the pupae hemo-lymphy of either G.melonnellan or H.armigera Hb could be used as the main component of the artificial diet for the development of T. evenecens, T. pintoi, T.chilonis. There was no significant difference in their parasitism, survival, percentage of pupation, adult emergence and reproduc-tively when the pupae hemolymphy of G.melonnella was used instead of that of H.armigera Hb [4; 5].
spp, (T. Evanescens, T. Pintoi, T. Chilonis) , 2017 year. (+ 26 ±2 ° C, RH 75 ± 3%)
№ Composition of medium% The amount of dam- age% Development degree of Trichogramma spp generation in artificial medium by days Genders proportion 8': ?
larva period Pupa period Mature period (imago) larva period
1. T/E Trichogramma evanescens 68.3 1.5 ± 0.3 4.3 ± 0.3 4.2 ± 0.5 3.5 ± 0.5 1:4
TEX TE2 TE3 TE 4
41.5 ±_0.5 15.5 ± 0.5 20.5 ± 0.5 22.5 ± 0.5
2. T/P Trichogramma pintoi 81.8 1.6 ± 0.3 4.6 ± 0.3 4.4 ± 0.5 4.2 ± 0.5 1:5
TP x TP 2 tp 3 TP 4
45.5 ±_0.5 13.5 ± 0.5 20.5 ± 0.5 20.5 ± 0.5
3. T/CH Trichogramma chilonis 83.5 1.8 ± 0.3 5.1 ± 0.3 4.8 ± 0.5 5.9 ± 0.5 1:7
TCj TC2 TC3 TC 4
45.2 ± 0.5 13.2 ± 0.5 21.6 ± 0.5 20.0 ±_0.5
Control 91.3 1.4 4.1 3.7 7.5 1:7
H.armigera eggs
It is obvious in the research, prepared all mediums of diets are harmed with Trichogramma spp generation and
put their eggs [4; 5; 6].
References:
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