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Yunuskhanov Shavkat, doctor of biological sciences, professor, chief of laboratory biochemical genetics, Institute of Genetics and plant Experimental Biology of the Academy of Sciences of the Republic of Uzbekistan
E-mail: [email protected] Abdurazakova Zumrad Lutfullaevna, Senior researcher, candidate of biological sciences, Institute of Genetics and plant Experimental Biology of the Academy of Sciences of the Republic of Uzbekistan
E-mail: [email protected]
Kurbanbaev Ilkham Djumanazarovich, doctor of biological sciences, Scientific secretary, Institute of Genetics and plant Experimental Biology of the Academy of Sciences of the Republic of Uzbekistan
E-mail: [email protected] Narimanov Abdujalil Abdusamatovich, doctor of agricultural sciences, professor, director, Institute of Genetics and plant Experimental Biology of the Academy of Sciences of the Republic of Uzbekistan
E-mail: [email protected] Azimov Abdulahat Abdujabborovich, Senior researcher, PhD., Institute of Genetics and plant Experimental Biology of the Academy of Sciences of the Republic of Uzbekistan
E-mail: [email protected]
PROTEIN MARKERS IN COTTON SPECIES G.HIRSUTUM L., G.BARBADENSE L. AND THEIR CORRELATION WITH VARIOUS CHARACTERS
Abstract. The data on the study of protein markers of cotton seeds of the species G.hirsutum L. and G.barbadense L. are presented. It is shown that the studied cotton species differ in the content of water soluble protein markers, designated according to their electrophoretic mobility by the symbols H-0.13 and B-0.18 and the buffer fraction, denoted by the characters AU and BD. Inheritance of protein markers H-0.13 and B-0.18 corresponds to monohybrid co-dominant cleavage. The loci of protein markers H-0.13 and B-0.18 are interconnected with a number of biochemical and morphological characters. Protein markers H-0.13 and B-0.18 turned out to be convenient when studying the genealogy of G.hirsutum L. and G.barbadense L. species. In diploid cotton species of the genomic group D, protein H-0.13 was found in seeds of the species G. trilobum, and protein B-0.18 - in the species G. thurberi, which suggests that these cotton species participated in the formation of the amphidiploid species G.hirsutum L. and G.barbadense L. The cotton species studied also differ in the composition of protein markers buffer fraction. The species of cotton G.hirsutum L. contains protein markers designated by the symbols AC, BD, and BCD, whereas in the species G.barbadense L. there are only protein markers AC. The cleavage of protein markers AC and BD in generations of interspecific hybrids depends on the combination of hybrids. In some F2 hybrids, the proteins AC and BD are inherited linked and correspond to monohybrid codominant cleavage, while in other F2 hybrids various combinations of these markers are formed.
Keywords: cotton, water-soluble proteins, buffer-soluble proteins, protein markers, G.hirsutum L., G.barbadense L., linkage group.
Introduction
Genetic markers identified on the basis of hereditary polymorphism of proteins, enzymes and DNA fragments proved to be powerful tools for identifying
the range of population and species variability, studying phylogenesis, and the degree of genetic similarity of various species. The development of these works led to the formation of a new direction in the field of fundamental and applied plant for studying genetic diversity, analyzing the genetic structure of the source genetics,
which was called marker-associated selection. With the introduction of molecular genetic methods in the selection process, new opportunities are opened material, identifying and controlling the genetic purity of genotypes. One of the ways to assess genotypes at the molecular level is the protein marker method based on the biological specificity of proteins detected by electrophoresis. The use of electrophoretic analysis of storage proteins makes it possible to evaluate genotypes by their genetic purity.
Materials and methods of research Seeds of various cotton varieties of the medium fiber species G.hirsutum L. and the fine fiber species G.barbadense L. of the Institute of Genetics and Experimental Plant Biology of the Academy of Sciences of the Republic of Uzbekistan were used as material for the study. Seed preparation for protein isolation and electrophoretic analysis was performed according to the methods described earlier by Yunuskhanov Sh. [1]. Result and discussion
Types of cotton G.hirsutum L. and G.barbadense L. have species-specific protein markers designated by symbols in the medium-fibrous species G.hirsutum L.- H-0.13 and in finegrained G.barbadense L. species - B-0.18, detectable in the electrophoretic composition of proteins of the water-soluble fraction of defatted seeds [2-4]. Different grades of these cotton species, with the exception of these protein markers, are almost identical in the main components of the electrophoretic spectrum of the water-soluble fraction. Molecular masses of protein markers H-0.13 and B-0.18, established by the method of sedimentation analysis, are 120 and 112 KD, respectively [5]. Inheritance of protein markers H-0.13 and B-0.18 in generations of interspecific hybrids corresponds to monohybrid codominant cleavage. However, in the study of the interspecific reciprocal hybrid of G. hirsutum L. and G. barbadense L. cotton obtained by crossing, where the species G. hirsutum L. was represented by dwarf plants obtained as a result of the mutation induced earlier by gamma irradiation [6], and the species G. barbadense L.- C-6037 strain an anomalous pattern was observed [7]. A radiomutant form characterized by dwarfism was isolated among the hybrids of cotton M2 obtained by pollination of flowers of plants of grade C-4727 with
pollen of plants of Tashkent-3 type ofmedium-fibrous species G.hirsutum L. irradiated with Co60 gamma rays at a dose of 1.0 kR [8]. The sign of dwarfism has been consistently split into normal plants: dwarf plants 1: 2. To study the relationship between the inheritance of species-specific marker proteins H-0.13 and B-0.18 with the sign of dwarfism of cotton plants, we obtained reciprocal hybrids between the radiomutant dwarf form of the medium fibrous species G.hirsutum L. and with the Izhod strain of the fine-fibered species G.barbadense L. However, neither the F1 plants nor the F2 hybrids showed a dwarfism sign, although the hybrid obtained with the participation of the Red-leaved Akala of the species G.hirsutum L. has a split ratio of 2: 1 (dwarf: tall plants). A similar picture was observed in the combination of the reciprocal hybrid, where one of the parents used the tall fine-fibrous variety C-6037 for crossing. Dwarf plants did not appear in the Fx and F2 populations of reciprocal hybrids. The hybrid obtained by crossing a dwarf form with the variety C-6037 of a thin-fibrous species of G. barbadense L. has been studied in various generations. The hybrid obtained by crossing a dwarf form with the variety C-6037 of a thin-fibrous species of G. barbadense L. has been studied in various generations. Plants of the dwarf form of the species G. hirsutum had a protein marker H-0.13, and in plants of the species G.barbadense L. the homologous marker was represented by protein B-0.18.
Plants of the species G. hirsutum had a protein marker H-0,13, and in plants of the type G.barbadense L. the homologous marker was represented by protein B-0.18. When analyzing phenotypic class relationships among generations of interspecies hybrids, including F2, F9 and F10, plants containing only one type of protein (H-0.13 or B-0.18) were considered as homozygotes for the corresponding gene, and plants containing both proteins (H-0.13 and B-0.18), as heterozygotes. A number of polymorphic progeny polymorphs showed a significant change in the ratio of the phenotypic classes, which manifested itself as a shift towards an increase or decrease in the proportion of a homozygous phenotypic class, or as a significant increase or decrease in the proportion of the heterozygous class. The electrophoretic composition of the seed proteins of the cotton samples under study remained unchanged up to the ninth generation of hybrids, but in the tenth generation some heterozygous plants produced offspring, in the seeds of which there was no protein with an electrophoretic mobility of 0.70. Such plants belonged to the homozygous phenotypic class, in which the marker H-0.13 is detected. In the work of M. A. Abzalov et al. [9], using this dwarf line in his studies on the relationship of traits, they came to the conclusion that this line contains the lethal Letdw gene, the heterozygous state of which determines the sign of dwarfism of plants, and the homozygous forms die in the early stages of seedling development. Consequently, the
data obtained by us on anomalous splitting can be explained, apparently, by the presence of a lethal gene in the mutant dwarf line of cotton. Investigation of seed proteins ofvarious interspecific hybrids has shown that the inheritance of protein markers H-0.13 and B-0,18 corresponds to monohybrid splitting and is interrelated with a number of economically valuable traits [4, 10-12]. Below are the clutch groups between protein markers H-0.13 and B-0.18 with a number ofbiochemical and morphological features of cotton:
PrH-0,13 _ prH-0,70 _ Fch_prH-0,23_ per0,33 _ PeelW — F^ua'ity — Flength —
-gweight _plantheight_Bollquantity;
PrB-0,18 _ prB-0,70 _ Fcb _prB-0,15 _Per0,33 _ PeelW _ F^uaHty _ Flength _
_ gweight _plantheight _ Bollquantity, where with dashes marked closely linked loci and without dashes _ recombined loci;
Pr _ Protein, the figure in the index is the electrophoretic mobility;
Fch and Fcb _ loci encoding fuzz on the chalazal part of seeds G.hirsutum L. and G.barbadense L.;
Per033 _ Peroxidase _ (peroxidase);
PeelW _ Peel weight _ (skin weight);
F qmii^ _ fiber quality;
F length _ fiber length;
Sweight _ seed weight-
Plant height _ plant height;
Boll quantity _ the number of boxes.
Protein markers H-0.13 and B-0.18 were also useful in studying the origin of amphidiploids G.hirsutum L., and G.barbadense L. [4]. It is assumed that cotton amphidiploids were formed by the fusion of the genomes of diploid species of the genomic group A (G.herbaceum) and the genomic group D (G.raimondii). When studying the protein spectrum of these groups of cotton, it turned out that representatives of cotton genomic group A do not contain protein markers H-0.13 and B-0.18, and representatives of cotton genome group D are divided by this trait into different groups. The type of cotton G.raimondii contains a protein with an electro-phoretic mobility close to H-0.13, but differs from it in other parameters. Protein protein marker H-0.13 was found in the species G.trilobum, in species G.thurberi _ B-0.18, other species of this genomic group simultaneously contain proteins identical in electrophoretic spectrum to proteins H-0.13 and B-0.18. Protein H-0.13 as in the species G.hirsutum L. and the species G.trilobum undergoes changes during their long-term storage. The electrophoretic mobility of the protein marker H-0.13 in long-stored seeds becomes identical with the protein B-0.18. Based on the data obtained, it can be assumed that the type of cotton G.trilobum from the genomic group D also participated in the formation of G.hirsutum L. and the type of G.thurberi in the formation of G.barbadense L.
Rf
I^SS
m ^-B
12 3 4
Figure 1. Electrophoregrams of water-soluble (1 and 2) and buffered soluble (3 and 4) proteins of defatted cotton seed flour. (1 and 3 - G. barbadense L., 2 and 4 - G.hirsutum L.)
The second group of protein markers was found in the composition of the buffer-soluble fraction of proteins of defatted seeds after removal of the water-soluble fraction [13]. This group of markers consists of two electrophoretic components, both in G.hirsutum L. species and in varieties of G.barbadense L. and according to their electrophoretic mobility are designated by the symbols A, B, C and D. Protein markers H-0.13 and B-0.18 are detected both in the water-soluble fraction of defatted seeds and in the buffer-soluble fraction, however, proteins A, B, C and D _ only in the buffer-soluble fraction (Fig. 1). The study of the buffer-soluble proteins of various varieties of cotton showed that G.hirsutum L. varieties in the content of components A, B, C and D are polymorphic and are divided into groups containing AC, BD and BD + C, while G.barbadense L. contain only one group of proteins _ AC. AC proteins in G.hirsutum L. species are found in Akala varieties and in varieties derived from them, as well as in the wild type cotton G.hirsutum L., ssp.mexicanum var. nervosum. The splitting of these proteins in generations of interspecies cotton hybrids depends on the combination of hybrids. There are combinations of hybrids in which the cleavage of the proteins AC and BD corresponds to normal monohybrid cleavage [13], with the loci encoding the proteins A and C on the one hand and B and D on the other are concatenated: F2 (AC x BD) = AC + 2(AC+BD) + BD In a number of combinations of hybrids in F2, an anomalous cleavage occurs, up to the formation of various combinations:
A A
AC BD AC BD
F,(AC xBD) =AC + (AC+BD) +BD +AD +AB + BC +A+ H-0,13 B-CUS +B+C+D
The G.hirsutum L. cotton varieties derived from hybridization between G.hirsutum L. and G.barbadense L. show only
protein AC markers. Figure 2. The interrelation of protein markers H-0,13,
The absence of protein markers of BD in the varieties of B-0,18 and buffer-soluble proteins A, B, C and D
fine-grained G.barbadense L. seems to indicate the incompat- Conclusion
ibility of loci of protein markers B-0.18 and protein markers The identified protein markers are promising for the bio-
BD: technological characteristics of the cotton gene pool and its use
in solving problems of plant breeding and seed production.
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