Applying polymerase chain reaction and fluorescence microscopy methods for Discrimination new self compatible almond genotypes (original) (raw)
Related papers
2016
Almond [Prunus amygdalus Batsch syn. Prunus dulcis (Mill.) D.A.Webb] trees are either self- or cross-incompatible, which results in lower fruit set and yields. Flower bagging, fluorescence microscopy, and polymerase chain reaction (PCR) were used to discriminate between selfcompatible genotypes obtained from crosses of the self-incompatible female parents (‘121’ and ‘4’) with the self-compatible male parent (‘Tuono’). This study was performed on 80 almond genotypes. The results of this study showed that, in the first cross (‘121’ × ‘Tuono’), genotypes 5, 11, 13, 14, 17, 20, 27, 29, 31, 35, and 38 were identified as being self-compatible and, in the second cross (‘4’ × ‘Tuono’), genotypes 1, 2, 10, 11, 12, 15, 21, 23, 25, 32, 37, 38,and 40 were found to be self-compatible. There were some promising genotypes based on self-compatibility and nut and kernel characteristics; for example, genotype 40 had the highest mean fruit and kernel weights at 2.9 and 1.3 g, respectively. PCR can be used to identify self-compatible genotypes at the juvenile stage. Flower bagging under favourable climatic conditions not only discriminated between self-compatible almond genotypes, but can also be used to measure fruit set percentages. Flower bagging and fluorescence microscopy can be used to determine the level of self-incompatibility. Fluorescence microscopy identified self-incompatible genotypes, even under unfavourable conditions. In general, a combination of all three methods is recommended to increase the accuracy of detecting selfcompatible genotypes of almond.
Detecting and determining self-incompatibility alleles in almond genotypes using molecular method
agriRxiv, 2019
One of the problems in almond production is self-incompatibility in this plant, which is considered as an important improvement point for this tree. Self-incompatibility causes non-uniformity and garden management problems. Most cultivars of almonds have gametophytic self-incompatibility that is controlled by a multi-allelic gene site. The inoculation inhibitor factor in this inhibitory system is the stop of pollen tube growth in the style. This study aims to detect and determine the self-compatible genotype from among the studied samples and determine the self-incompatibility alleles in the studied masses. For the experiment, the leaf samples were collected from 100 almond genotypes that had good products in recent years. The DNA of young leaf samples in these genotypes was extracted using Gept and Celeg (1989) method with a few changes. Today, various methods have been invented for detecting the genotypes and self-compatible cultivars from selfincompatible cultivars as well as S a...
Almond Breeding to obtain new Self -compatible Genotypes by Molecular Method
2016
Almond tree has tolerance to drought and its fruit has high nutritional value. In almond production, self –incompatibility is one of the major problems and to solve this, many breeding programs have been conducted around the world. In this study we used a couple of primers for identification and determination of S-alleles in self-compatible and selfincompatible almond genotypes. Also in this research We studied some wild genotypes to evaluate their S-allels .The results showed , using allele specific primers SfF-SfR, Cebador 2-Cebador8 and ConF-ConR ,we obtained respectively 450bp,1200bpand 1200bp bands in self-compatible genotypes .wear as using of AS1II-AmyC5R primers were produced 1100,1200,800 and 600 bp bands for S1, Sf/S3, S2 and S5/S25 respectively. As well as using two primers (Cebador1Cebador3) produced a band related to S1 allele and using S3F-S3R2 we could recognized between S3 and Sf bands produced by As1II and AmyC5R pair primers. The results showed S2F and S2R primers ...
2012
Self-incompatibility in almond and Prunus species is an important trait that prevents self-fertilization. Selfincompatibility in almond is controlled gametophytically by the multiallelic S-locus. Present study was done in order to identification and preliminary selection of self compatible progenies resulted from controlled crosses in almond using specific primer SfF/SfR. Some important morphological traits of parental crosses were evaluated using almond descriptor. Also, progenies (F1) of five crosses inculding; A (Tuono × 101 Genotype), B (Supernova × 101 Genotype), C (Genco × Shahrood 21), D (Tuono × Shahrood 12) and E (Tuono × shahrood 17) were tested using PCR in order to DNA amplification and self-compatibility evaluation. The result of PCR method found that using SfF/SfR pair primer, self-compatible progenies showed 449 bp bands, while this band not observed in self-incompatible progenies. In addition, all self- incompatible progenies appeared no S1 allele at any condition. A...
2017
The evaluation of an almond collection using morphological variables and identification of self-incompatibility genotype is useful for selecting pollinizers and for the design of crossing in almond breeding programs. In this study, important morphological traits and self-incompatibilities in 71 almond cultivars and genotypes were studied. Simple and multiplex specific PCR analyses were used in order to identify self-incompatibility alleles. Based on the results, cultivars and genotypes including ‘Dir Ras–e-Savojbolagh’, ‘D-124’, ‘D-99’, ‘Shahrood 12’, ‘Tuono’, ‘Nonpareil’, ‘Price’, ‘Mirpanj-e-Tehran’, ‘Pakotahe-e- Taleghan’, ‘V-13-34’, ‘V-16-8, ‘V-11-10’, ‘Zarghan 10’, ‘Uromiyeh 68’, ‘Barg dorosht-e-Hamedan’ and ‘Yazd 60’ were late flowering and had the highest quality of nut and kernel characters. The result of the PCR method using combined primers AS1II and AmyC5R showed amplification of ten self-incompatibility alleles (S1, S2, S3, S5, S6, S7, S8, S10, S12,and S unknown allele) ...
Identification of Self-Incompatibility Alleles in Almond and Related Prunus Species Using PCR
Acta horticulturae, 2003
The almond, Prunus dulcis Miller which belongs to Rosaceae family, is one of the most important commercial and oldest cultivated tree nut crops. Almonds are classified as a 'nut' in which the edible seed is the commercial product. Therefore, pollination and fertilization are necessary in almond. The characteristic of cultivated almond to express gametophytic self-incompatibility discourages self-fertilization and favors cross pollination. Genetic control of pollen-pistil self-incompatibility is through a single gene (S) which exists in a series of alleles S 1 to S x. Compatibility of pollen-pistil in almond is an important consideration in planning crosses in breeding program and in choosing pollinizers for orchard planting. Identification of self-(in) compatibility in almond carried out by molecular and controlled pollination methods. In this study, identification of S-alleles in 37 Iranian almond cultivars and genotypes was carried out by PCR method with using degenerate primers of EM-PC3consRDEMPC2cons FD, PaconsI-Fand EM-PC1consRD. In this way the size of S-alleles were estimated based on bands which amplified with second intron. The results confirmed self-incompatibility in cultivars and most genotypes. However, the S f-like allele (in size) was observed in A 9 and A 36 genotypes. If these results are confirmed by sequencing the S f allele, it will be first time to identify self-compatible genotype in Iranian almond genotypes.
Morphological and Molecular Characterization of a New Self-Compatible Almond Variety
Agriculture
Almonds are one of the most popular nuts, cultivated in countries with Mediterranean climates. In an almond orchard of the self-incompatible cultivar ‘Ferragnes’ in Greece, a tree with different morphological characteristics and signs of self-compatibility was observed. The aim of this study was to study the phenotype, investigate the self-compatibility trait, and elucidate the phylogenetic background of this tree, named ‘Mars’. Morphological traits and kernel and nut characteristics were measured in ‘Mars’, ‘Ferragnes’, ‘Tuono’, and ‘Lauranne’ cultivars. The self-compatibility trait of almonds is attributed to the Sf allele; thus, its existence was investigated in ‘Mars’ by PCR amplification. Moreover, the S-RNase genes of all the cultivars were sequenced. The genetic profile of ‘Mars’ was identified using eight SSR molecular markers and compared with the ‘Ferragnes’, ‘Ferraduel’, ‘Texas’, ‘Tuono’, and ‘Lauranne’ cultivars. The morphological traits suggest that ‘Mars’ is more simil...
2015
Self-incompatibility is one of the most important difficulties in almond production which reduce fruit set dramatically and makes orchard management difficult. Therefore, breeding almond to produce self-compatible genotypes is very important. In this research identification and screening of 86 almond progenies obtained from selfing Touno after the selfpollination by PCR reaction with specific primers of CEBASf and AS1. PCR results confirmed the situation of selfcompatible hybrids. In addition, it indicated that, frequencies of Sf, and S1 was 100% and 50% in progenies respectively. Self-compatible hybrids had been identified that can be used in almond breeding programs particularly to development the monoculture of almond orchards. So to identify and screening homozygous self-compatibility almonds be capable of be another step towards creating monoculture of almond and use in breeding programs further.
Identification of self-incompatibility genotypes of almond by allele-specific PCR analysis
TAG Theoretical and Applied Genetics, 2000
In almond, gametophytic self-incompatibility is controlled by a single multiallelic locus (S-locus). In styles, the products of S-alleles are ribonucleases, the S-RNases. Cultivated almond in California have four predominant S-alleles (S a , S b , S c , S d ). We previously reported the cDNA cloning of three of these alleles, namely S b , S c and S d . In this paper we report the cloning and DNA sequence analysis of the S a allele. The S a -RNase displays approximately 55% similarity at the amino-acid level with other almond S-RNases (S b , S c , and S d ) and this similarity was lower than that observed among the S b , S c and S d -RNases. Using the cDNA sequence, a PCR-based identification system using genomic DNA was developed for each of the S-RNase alleles. Five almond cultivars with known self-incompatibility (SI) geno-types were analyzed. Common sequences among four S-alleles were used to create four primers, which, when used as sets, amplify DNA bands of unique size that corresponded to each of the four almond S-alleles; S a (602 bp), S b (1083 bp), S c (221 bp) and S d (343 bp). All PCR products obtained from genomic DNA isolated from the five almond cultivars were cloned and their DNA sequence obtained. The nucleotide sequence of these genomic DNA fragments matched the corresponding S-allele cDNA sequence in every case. The amplified products obtained for the S a -and S b -alleles were both longer than that expected for the coding region, revealing the presence of an intron of 84 bp in the S a -allele and 556 bp in the S b -allele. Both introns are present within the site of the hypervariable region common in S-RNases from the Rosaceae family and which may be important for S specificity. The exon portions of the genomic DNA sequences were completely consistent with the cDNA sequence of the corresponding S-allele. A useful application of these primers would be to identify the S-genotype of progeny in a breeding program, new varieties in an almond nursery, or new grower selections at the seedling stage.
Self and cross incompatibility traits analysis in some diseases tolerant almond genotypes by PCR
African Journal of Microbiology Research, 2012
Almond is one of the oldest nut crops in the world and has a main problem called self-incompatibility. Identification of S alleles in almond cultivars and genotypes is essential for breeders and growers. The aim of this study was to identify S alleles pattern in 30 late bloom and resistant to aphid, mite, Monilia laxa and "Polystigma occharaceum" fungus almond genotypes from different geographical regions of East-Azerbaijan Iran. Most of the S alleles were amplified using Pru-C 2 /Pru-C 5 R primer pair and size of alleles ranged between 2000-220 bp in genotypes. Considering all of the S alleles amplified using 5 primer pairs in thirty numbers of the genotypes two S alleles were detected and showed their fully selfincompatibility pattern, in 10 genotypes only one of the S alleles were.