Оцінка стійкості ліній перцю солодкого (Capsicum Annuum L.) до фузаріозного в’янення на рівні культури in vitro та in vivo

С. І. Кондратенко, О. Ю. Гарт, О. В. Черненко

Анотація


ASSESSMENT OF SWEET PEPPER LINES (CAPSICUM ANNUUM L.) TO FUSARIUM WILT AT THE LEVEL OF CULTURE IN VITRO AND IN VIVO

S. I. Kondratenko, O. Yu. Gart, O. V. Chernenko

 

Introduction. In the last years one of the most harmful diseases of sweet pepper in Ukraine is the disease of wilt, the main causal organisms of which under conditions of the Forest-Steppe zone is the complex of pathogenic fungi of the Fusarium Link genus. The investigations of scientists proved that the resistance to this disease is controlled genetically and the intensity of its virulent properties relative to the object of investigations depends on the ontogenesis phase of a plant and directions of breeding process conducting. Until today genetic physiologic mechanism of sweet pepper plants resistance to fusarium wilt are studied still extremely insufficiently. Effective informative criterion for selection of resistance to fusarium wilt genotypes of sweet pepper is the use of the method of cell selection in vitro, which involves the study in a sterile culture the reaction of somatic and meristem cells in the nutrient medium with addition of the filtrate of cultural liquid (FIL) of the main pathogens of the disease.

Aim of the investigation is ascertainment the level of manifestation of the trait of sweet pepper resistance to fusarium wilt at the level of sporophyte generation in culture in vitro, elucidation correlative connections between the main quantitative traits of a linear material under conditions of the natural provocative background (culture in vivo).

Methods. The object of investigation were 10 breeding-valuable lines of sweet pepper of the Institute of Vegetable and Melons of NAAS breeding. The standard was the adapted domestic variety of Pioneer. Lines were grown under conditions of provocative stationary background (film greenhouse) during 2010-2016 years. Evaluation of lines by their resistance to fusarium wilt and the complex of different economic traits was conducted according to the European classifier. In the investigations there was studied the reaction of callus cells obtained from primary explants isolated from hypocotyls and cotyledons against the background of presence in the nutrient medium MS (Murashige and Skoog, 1962) selective agent for selection of valuable genotypes – 30-50 % FIL of the complex of phytopathogenic fungi of Fusarium sp. sp. genus. The volume of received calluses was counted on the 24th day of cultivation. The MS nutrient medium without content of FIL was the control.

Results and Discussion. As it was shown by the results of experiments the volume of cells callus growth directly depended on such factors: level of resistance of the specific sweet pepper genotype; origin of primary explants introduced into the culture in vitro; concentration of FIL in the content of nutrient medium. It was revealed that in culture in vitro the greatest inhibition of callus growth was recorded when in the content of nutrient medium 30 % FIL was used. For today it provided us the possibility to use just this concentration to work out the method for selecting valuable initial breeding forms of sweet pepper in culture in vitro. First of all exactly in the culture of hypocotyls explants the variant of the experiment with adding of 30 % FIL to the nutrient medium proves that they have the most contrast (selective) reaction of callus cells of the tested sweet pepper genotypes by their resistance to fusarium wilt. In contrast to this, the use of 50 % solution of FIL in nutrient medium was marked by active increase in the intensity of callus growth relative to the control. Under conditions of stationary provocative background (perennial permanent one-crop system of Solanaceae) in a film greenhouse during 2013-2016 years there is determined the reaction level of the mentioned trait working collection of sweet pepper genotypes by the degree index of their affection with fusarium wilt according to the modified grading scale of evaluation. Generalization of data makes it possible to state the fact that statistically reliably (at the level p ≥ 0,05) researched samples by this trait are distributed as follows: samples UL0500371, UL0500387, UL0500391 were stable in the grade 7 of PEB scale (up to 10.1 % of affected plants), samples UL0500388, UL0500386, UL0500374, UL0500389, UL0500391 were stable in the grade 5 (up to 35.1 % of affected plants), the sample, UL0500375 and the variety-standard Pioneer (Ul0500001) were stable in the grades 3 and 1.

The correlation analysis has been made between quantitative and biometric indices of plants and fruits of sweet pepper lines. In all 19 quantitative traits were involved in the correlation matrix statistically reliable (at the level p ≥ 0,05) turned out correlation connection in 11 couples of traits. It should be especially noted the inverse correlation connection between the growth of callus obtained from hypocotyls explants and the trait of the affection degree of a sample by fusarium wilt that makes it possible to use this trait  as a criterion for selecting resistant genotypes of sweet pepper and to optimize the methods for breeding process conducting. The established direct correction connections between such sample parameters as “Thickness of pericarp”, “Fruits quantity on a plant” and “Degree of affection by fusarium wilt” give us the possibility to forecast the level of initial material resistance to this disease.

It is determined that at the level of sporophyte progeny the investigated samples varied by different traits of valuable genetic lines (National Centre for Plant Genetic Resources of Ukraine, collection № 77) by their resistance to fusarium wilt under conditions of natural provocative background from 4.3 to 54.8 % (border grades of resistance are from 7 to 1).

It is ascertained that biotechnological methods of cell selection in vitro make it possible to single out for subsequent breeding researches at early of the progress valuable by the complex of traits initial material of sweet pepper. Obtained results of the correlation analysis confirmed the important theoretical and practical significance and constructiveness in evaluating and further selecting valuable by the complex of traits initial breeding material with the aim of further obtainment of competitive native varieties and hybrids F1 of sweet pepper which will make it possible to join traits of high productivity, content of valuable biochemical components and resistance to diseases in their genotypes.

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