Potential of trispecies bridge crosses and random amplified polymorphic DNA markers for introgression of Medicago daghestanica and M. pironae germplasm into alfalfa (M. sativa)

Genome ◽  
1993 ◽  
Vol 36 (3) ◽  
pp. 594-601 ◽  
Author(s):  
T. J. McCoy ◽  
C. S. Echt

This report describes the production and cytology of the first interspecific hybrids between cultivated alfalfa (Medicago sativa L.) at the diploid level (2n = 2x = 16) and the diploid (2n = 2x = 16) perennial species M. daghestanica and M. pironae. An ovule–embryo culture technique was required to rescue hybrid embryos and all hybrids were diploid. Predominately bivalent chromosome pairing was observed at meiotic metaphase. All F1 hybrids were male and female sterile and no species backcross progeny could be produced. We discovered that trispecies hybrids could be efficiently recovered via crossing diploid F1 interspecific hybrids of M. sativa × M. rupestris with either M. daghestanica or M. pironae. Ovule–embryo culture was also required to recover these trispecies hybrids with recovery efficiency of trispecies hybrids about 10 times greater than for bispecies hybrids. Most chromosomes paired as bivalents in the trispecies hybrids. Importantly, progeny can be recovered from crossing the trispecies hybrids with M. sativa. Therefore, the M. sativa × M. rupestris hybrids provide a bridge cross to potential introgression of M. daghestanica or M. pironae germplasm. Analysis of randomly amplified polymorphic DNA (RAPD) markers in the trispecies hybrids indicates that RAPD markers offer considerable potential for assaying germplasm introgression following complex hybridizations of the type reported here.Key words: randomly amplified polymorphic DNA, Medicago interspecific hybrids, embryo rescue.

Jurnal Biota ◽  
2021 ◽  
Vol 7 (1) ◽  
pp. 42-50
Author(s):  
Muhammad Khoerol Anam ◽  
Adi Amurwanto ◽  
Kusbiyanto Kusbiyanto ◽  
Hendro Pramono ◽  
M Husein Sastranegara ◽  
...  

Segara Anakan areas can be divided into three different regions according to their salinity. Salinity differences suggested that Commerson’s anchovy population in that area can be divided into three subpopulations due to genetic differences. Genetic differences among subpopulation can be assessed through a population genetic study using random amplified polymorphic DNA. This study aims to evaluate the genetic variation and differences of Commerson's anchovy (Stolephorus commersonnii) collected at three different water salinities in Segara Anakan estuary Cilacap Indonesia. Total genomic DNA was isolated using the Chelex method. Genetic diversity and differences were assessed using RAPD markers and were analyzed statistically using an analysis of molecular variance, as implemented in Arlequin software.  The results showed that high genetic diversity was observed within the subpopulations. However, no significant genetic differences were observed among subpopulations which indicate genetic similarity. A high number of offspring are likely to cause high genetic variation within subpopulations.  Adult and larvae migration is the cause of genetics similarity across Segara Anakan. Another impressive result is that water salinity did not affect the genetic characteristic of Commerson,s anchovy. Genetic similarity of Commerson’s anchovy indicates that Segara Anakan forms a single genetic conservation unit.


Genome ◽  
1989 ◽  
Vol 32 (1) ◽  
pp. 6-11 ◽  
Author(s):  
T. J. McCoy ◽  
G. L. Quarisa

Diploid (2n = 2x = 16), triploid (2n = 3x = 24), and tetraploid (2n = 4x = 32) interspecific hybrids between alfalfa (Medicago sativa L.) and M. papillosa Boiss. were recovered either from seed (the triploid hybrids) or from ovule–embryo culture (the diploid and tetraploid hybrids). Cytogenetic analysis of diploid interspecific hybrids (with one genome of M. sativa, designated S, and one genome of M. papillosa, designated P), indicated significant genomic affinity, with an average of 7.6 bivalents and 0.8 univalents per pollen mother cell. In contrast, cytogenetic analysis of the triploid interspecific hybrids (with one S genome and two P genomes) indicated little if any genomic affinity between M. sativa and M. papillosa. In 7 of 14 triploid hybrids analyzed no trivalent configurations were observed, and in the other hybrids, trivalent frequency ranged from 0.1 to 0.4 per pollen mother cell. Tetraploid interspecific hybrids with two S and two P genomes had predominantly bivalent pairing. Based on the lack of homology of S and P genomes, the tetraploid hybrids are basically allotetraploids (SSPP). Therefore, backcross progeny from crossing the tetraploid hybrids with tetraploid M. sativa have the genomic constitution SSSP. Univalents and trivalents were observed in first backcross (BC1) progeny, as expected, based on an allotetraploid interpretation. Most of the BC1 progeny were partially or completely male sterile, and female fertility was significantly reduced. Potential uses of homoeologous genomes such as M. papillosa in alfalfa genetic and breeding studies are discussed.Key words: cytogenetics, interspecific hybrids, ovule –embryo culture.


1985 ◽  
Vol 27 (2) ◽  
pp. 238-245 ◽  
Author(s):  
T. J. McCoy

An ovule–embryo culture method was used to produce the first interspecific hybrids between alfalfa (Medicago sativa L.) and Medicago rupestris M. B. Culture of fertilized ovules from the cross diploid (2n = 2x = 16) M. sativa (jpjp) × diploid (2n = 2x = 16) M. rupestris began 14 days after pollination. After 5 days in culture, the interspecific hybrid embryo was removed and transferred to fresh medium, where development into a plant occurred. Forty-six M. sativa – M. rupestris F1 hybrids have been recovered using this technique. All but one of the 46 F1 hybrids were diploid (2n = 2x = 16); the only exception was tetraploid (2n = 4x = 32). The most frequent meiotic configurations observed in the F1 hybrid plants were eight bivalents or seven bivalents and two univalents, indicating significant homology between M. sativa and M. rupestris genomes. However, pollen stainability (0–12%) and pollen germination (0–6%) were extremely low. Similar to the production of the F1, no first backcross (BC1) plants were obtained from seed; however, the ovule–embryo culture method was found to be a very effective method for recovering BC1 plants and hundreds of BC1 plants have been produced. The BC1 plants from crossing the F1 with diploid M. sativa were predominantly diploid. Medicago rupestris can now be considered a potential germplasm source for alfalfa improvement. The ovule–embryo culture method represents the first successful recovery of Medicago interspecific hybrids via some form of embryo rescue. Importantly, it appears this technique can be applied to other interspecific hybrid combinations in the Medicago genus.Key words: Medicago, alfalfa, embryo culture, interspecific hybrid.


2021 ◽  
Author(s):  
Mohammad Akmal

Somatic embryogenesis is the best demonstration of totipotency in higher plants in which somatic cell produce whole plant like zygotic embryo. It is also demonstrated that immature, weak, hybrid or sometimes inviable embryos can be saved through in vitro culture to prevents its degradation. It may help to cross the reproductive barriers when interspecific hybrids developed. Brasssica is an economically valuable oil yielding and vegetable crop and India is the largest producer of oil seed rape in the world. Various factors affect the embryo rescue in Brassica like growth stage of the embryos, types and composition of the rescue medium etc. The embryo regeneration potential can improve through the modification of culture conditions in both zygotic as well as somatic embryo. Except the embryo culture other parts like ovule, ovary culture can also be done to developed interspecific hybrids. This chapter is focused on the embryo rescue techniques in the genus Brassica and summarizes possible ways of improving the technique used.


Genome ◽  
1987 ◽  
Vol 29 (6) ◽  
pp. 853-858 ◽  
Author(s):  
C. R. Denton ◽  
T. J. McCoy

Interspecific hybrids between diploid (2n = 2x = 16) Medicago sativa L. and diploid (2n = 2x = 16) M. rhodopea Velen., were recovered using an ovule–embryo culture methodology. Most hybrids were vigorous, and morphological comparisons demonstrated that F1 hybrids were generally intermediate between that of the parents. Peroxidase isozyme phenotypes of the F1 hybrids confirmed hybridity. The chromosome number of most of the hybrids was diploid (2n = 2x = 16), with the exceptions of two triploids (2n = 3x = 24) and two tetraploid (2n = 4x = 32) plants. Chromosome pairing configurations in diploids were almost exclusively eight bivalents or seven bivalents and two univalents, indicating a high level of homology between the M. sativa and M. rhodopea genomes. However, the one triploid hybrid analyzed had only 0.4 trivalents per microspore mother cell indicating preferential pairing of parental genomes. Pollen stainability, pollen germination, and fertility of the diploid F1 hybrid plants were very low; however, it was possible to obtain backcross progeny (BC1) from seed. Pollen stainability, pollen germination, and fertility of the BC1 plants were also very low; however, most BC1 plants had workable levels of male and female fertility. The utilization of M. rhodopea in studies of the evolution of hexaploid Medicago species is discussed. Key words: interspecific hybrids, ovule–embryo culture, isozymes, Medicago.


HortScience ◽  
1999 ◽  
Vol 34 (7) ◽  
pp. 1256-1258 ◽  
Author(s):  
Feiga Gutman ◽  
Avinoam Nerd ◽  
Yosef Mizrahi ◽  
Dudy Bar-Zvi ◽  
Dina Raveh

Twenty-four genotypes of marula (Sclerocarya birrea subsp. caffra) were characterized using randomly amplified polymorphic DNA (RAPD) analysis. A distinct band pattern was obtained for each of the trees, using as few as four arbitrary 10-mer primers. Trees propagated vegetatively by grafting showed identical fingerprints. These results suggest that RAPD markers provide a useful system for documenting the identity of marula genotypes.


2008 ◽  
Vol 13 (1-2) ◽  
pp. 151 ◽  
Author(s):  
E. KIVIHARJU ◽  
J. LAURILA ◽  
M. LEHTONEN

A study was carried out to improve anther culture ability of the non-responsive cultivated oat, Avena sativa L. cv. Puhti by introgressing favourable alleles from the responsive wild red oat, Avena sterilis L. acc. CAV 2648. Anther culture ability of these parental lines and F2 progenies of their cross and two backcrosses was tested. Genotype effects were significant on all anther culture traits measured. The number of anther culture derived embryo-like structures was highest in acc. CAV 2648, and the number of green regenerants from the Puhti × CAV 2648 progeny. Anther culture response was greatly reduced in backcross progeny and was least in cv. Puhti. Random amplified polymorphic DNA (RAPD) was used to test for marker associations with oat anther culture traits in a population of 38 F2 progenies. Two RAPD markers were putatively associated with improved production of green regenerants (one derived from acc. CAV 2648 and the other from cv. Puhti). One marker putatively associated with decreased albino plant regeneration (derived from acc. CAV 2648). These markers might be useful for selecting alleles for better anther culture ability among progeny of planned crosses. In addition, three markers, derived from acc. CAV 2648, were putatively associated with decreased anther culture response rates.;


1995 ◽  
Vol 120 (5) ◽  
pp. 726-729 ◽  
Author(s):  
C.M. Ronning ◽  
R.J. Schnell ◽  
S. Gazit

The native American genus Annona contains many species that are cultivated for their edible fruit, including the custard apple (A. reticuluta L.), soursop (A. muricata L.), cherimoya (A. cherimola L.), sugar apple (A. squamosa L.), and interspecific hybrids, the atemoyas. RAPD analysis of A. cherimola. `Campa' and `Jete,' A. squamosa `Lessard,' and the atemoyas `Ubranitzki,' `Malali,' and `Kaspi' resulted in very distinctive patterns, indicating that RAPD markers, may be an efficient method of fingerprinting genotypes within and between Annona species. All 15 primers used generated repeatable, polymorphic patterns. An F1 population of `Jete' × `Lessard' was analyzed to determine the inheritance of the RAPD banding patterns. Fifty-two polymorphic loci were identified, which segregated in an expected Mendelian fashion.


2017 ◽  
Vol 7 ◽  
Author(s):  
Santosh P ◽  
Suresh B. Arakera

<p><strong>Polymerase chain reaction based random amplified polymorphic DNA (PCR-RAPD) markers were employed to assess genetic diversity in eight <em>Garcinia  cambogia</em>  genotypes. Among the 20 random primers used in the present investigation, 9 primers showed polymorphism. A total number of 227 bands were obtained from 9 primers, out of which 225 were polymorphic, showing 99.11% polymorphism. An average of 25.22 bands per primer was scored and average number of polymorphic bands found to be 25. The eight accessions fall into two major clusters. Cluster analysis showed that the red and yellow accessions cannot be regarded as two different varieties. The use of red and yellow fruits for commercial and medicinal purposes, respectively, is purely based on consumer preference. </strong></p>


2017 ◽  
Vol 23 (2) ◽  
Author(s):  
SUNITA BORDE ◽  
ASAWARI FARTADE ◽  
AMOL THOSAR ◽  
RAHUL KHAWAL

Ptychobothridean genera like Senga and Circumoncobothrium are the common parasites of fresh water fishes. The genotypic study of these parasites was taken by RAPD. The RAPD profile of these two parasites were not similar to each other as depicted by the band pattern in picture. These results suggest the presence of inter-specific polymorphism among cestode parasites of two different genera for RAPD analysis. The present study demonstrated that genetic differentiation of cestode parasites could be accomplished on the basis of genomic variation with polymorphic band pattern using RAPD. All the detected bands (PCR product) were polymorphic and band size ranged from 500-5000 bp in length. The RAPD of profiles using GBO-31, GBO-32, GBO-33, GBO-34, GBO-35 and GBO-36. Primers were able to characterize inter-specific polymorphism among the two genus ( Senga and Circumoncobothrium ). Genetic analysis suggests that Senga and Circumoncobothrium show genetic diversity with respect to RAPD patterns using all the six primers used for the present study. The genetic distance between the analyzed genuses ranged from 0.14 to 0.80. The differentiation of the two parasites on the basis of genetic markers could greatly facilitate study on the biology of these parasites.


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