Physiological and Molecular Studies on Some Rice Cultivars
Al Shymaa Zaky Al Mokadem;
Abstract
The present investigation was conducted to develop molecular genetic fingerprints for nine local rice cultivars, to elucidate relationships among these cultivars and to investigate the molecular basis of salt tolerance in nine rice cultivars.
Identification of nine rice cultivars
Identification based on SDS-PAGE protein electrophoresis
Data of SDS-PAGE for total protein in leaves revealed that bands with different molecular weights (MW) were detected and ranging from about 9.9 KDa to 166 KDa. The total number of bands among genotypes ranged from 27 in Sakha101 to 29 in Sakha104, Sakha103 and Giza178. The level of polymorphism was 38.5% among the studied cultivars.
Identification based on RAPD
Analysis of ten RAPD primers successfully produced multiple band profiles with a polymorphism percentage ranged from 30% to 81.82%. Primers produced band numbers ranging from 11 (primer OP- E19) to 23 (primer OP-A16) across cultivars the total number of reproducible fragments amplified by the ten primers reached 157 from which 74 were polymorphic fragments. This represented a level of polymorphism of 47.13% among the studied lines.
Identification based on SSR
Data of the amplified fragments using five SSR primers for the nine rice cultivars revealed that all primers succeeded in amplifying DNA fragments. Number of alleles in all tested primers is one allele per locus. The overall size of amplified products ranged from 137 bp (RM246) to 338 bp (RM166). There is no unique alleles were observed at all of the 5 tested SSR loci. The frequency of all alleles was 100% while the polymorphism was zero.
Phylogenetic Relationships
The phylogenetic dendrogram based on the overall markers (SDS-PAGE, RAPD and SSR- PCR) indicated two main groups: The first main group was subdivided into two subgroups. The first subgroup includes Sakha102 and each of Sakha105, Sakha103 and Sakha101. The second subgroup group was includes AgamiM5 and Sakha104. The second main group included Giza 178 and each of Giza 177 and Nabatat asmar. The similarity coefficient reached a highest value of 1.0 between Sakha102 and Sakha103, and reached a minimum value 0.0 between Giza 178 and Sakha105.
After conducting a series of laboratory experiments, greenhouse pot and application under saline field experiments so as to assess the role of exogenously applied validamycin A in inducing salt tolerance in two rice cultivars through enhancing the accumulation of trehalose, following conclusions can be drawn:
1- Salt stress-induced reduction in growth of rice plants was alleviated by exogenously applied validamycin A.
2- Improvement in growth of salt stressed plants of both rice cultivars was associated with increased photosynthetic rate.
3- Improvement in photosynthetic efficiency was partially associated with amount of photosynthetic pigments, probably due to osmo-protective effects of trehalose on chloroplast (thylakoid membranes).
4- Salinity stress decreased starch and total soluble protein contents concomitantly with increasing total soluble sugars, sucrose, trehalose, proline and free amino acids in both cultivars. Raising trehalose level in the plant tissues treated with validamycin A was accompanied by increase in the starch and protein content and appearance of new protein pattern of both cultivars.
5- Trehalose accumulation can enhance tolerance of both rice seedlings to salt stress by decreasing the concentration of MDA and H2O2 and increasing the transcript levels and activities of SOD, APX, CAT, GR, POD and PPO, and the contents of cartenoids, ascorbic acid, flavonoids, and tocopherols which, in turn, enhance the antioxidant ability and protect rice seedlings against oxidative stress induced by salt stress.
6- Production of antioxidants and membrane protecting compounds help plants accumulated trehalose to withstand salt exposure. It is reasonable to believe that specific membrane lipid classes and sustainable transport proteins are constitutive or induced under saline conditions which might contribute to salt tolerance acquired by trehalose accumulation.
7- Exogenous application of validamycin A caused high accumulation of K+, N and P and lowering the content of Na+ in both cultivars under salt stress conditions.
Further investigations are needed to validate the effectiveness of this antibiotic manipulation under field conditions and to establish the effective doses based on different salt stress levels.
1- Salt stress-induced reduction in number of seeds, grain yield per plant, 1000-grain weight and filled-grain percentage for both rice cultivars was alleviated by exogenous application of validamycin A as pre-sowing seed treatment under saline field conditions
2- Exogenous application of validamycin A was found to be most effective at AgamiM5 cultivar as compared to the other cultivar Skha103.
At the molecular level, alterations in protein pattern of the treated rice tissues revealed that validamycin A treatments induced the appearance of new set of protein bands concomitant with the disappearance of others. Moreover increment in TPS gene expression of both rice cultivars tissues over those untreated tissues was recorded.
Identification of nine rice cultivars
Identification based on SDS-PAGE protein electrophoresis
Data of SDS-PAGE for total protein in leaves revealed that bands with different molecular weights (MW) were detected and ranging from about 9.9 KDa to 166 KDa. The total number of bands among genotypes ranged from 27 in Sakha101 to 29 in Sakha104, Sakha103 and Giza178. The level of polymorphism was 38.5% among the studied cultivars.
Identification based on RAPD
Analysis of ten RAPD primers successfully produced multiple band profiles with a polymorphism percentage ranged from 30% to 81.82%. Primers produced band numbers ranging from 11 (primer OP- E19) to 23 (primer OP-A16) across cultivars the total number of reproducible fragments amplified by the ten primers reached 157 from which 74 were polymorphic fragments. This represented a level of polymorphism of 47.13% among the studied lines.
Identification based on SSR
Data of the amplified fragments using five SSR primers for the nine rice cultivars revealed that all primers succeeded in amplifying DNA fragments. Number of alleles in all tested primers is one allele per locus. The overall size of amplified products ranged from 137 bp (RM246) to 338 bp (RM166). There is no unique alleles were observed at all of the 5 tested SSR loci. The frequency of all alleles was 100% while the polymorphism was zero.
Phylogenetic Relationships
The phylogenetic dendrogram based on the overall markers (SDS-PAGE, RAPD and SSR- PCR) indicated two main groups: The first main group was subdivided into two subgroups. The first subgroup includes Sakha102 and each of Sakha105, Sakha103 and Sakha101. The second subgroup group was includes AgamiM5 and Sakha104. The second main group included Giza 178 and each of Giza 177 and Nabatat asmar. The similarity coefficient reached a highest value of 1.0 between Sakha102 and Sakha103, and reached a minimum value 0.0 between Giza 178 and Sakha105.
After conducting a series of laboratory experiments, greenhouse pot and application under saline field experiments so as to assess the role of exogenously applied validamycin A in inducing salt tolerance in two rice cultivars through enhancing the accumulation of trehalose, following conclusions can be drawn:
1- Salt stress-induced reduction in growth of rice plants was alleviated by exogenously applied validamycin A.
2- Improvement in growth of salt stressed plants of both rice cultivars was associated with increased photosynthetic rate.
3- Improvement in photosynthetic efficiency was partially associated with amount of photosynthetic pigments, probably due to osmo-protective effects of trehalose on chloroplast (thylakoid membranes).
4- Salinity stress decreased starch and total soluble protein contents concomitantly with increasing total soluble sugars, sucrose, trehalose, proline and free amino acids in both cultivars. Raising trehalose level in the plant tissues treated with validamycin A was accompanied by increase in the starch and protein content and appearance of new protein pattern of both cultivars.
5- Trehalose accumulation can enhance tolerance of both rice seedlings to salt stress by decreasing the concentration of MDA and H2O2 and increasing the transcript levels and activities of SOD, APX, CAT, GR, POD and PPO, and the contents of cartenoids, ascorbic acid, flavonoids, and tocopherols which, in turn, enhance the antioxidant ability and protect rice seedlings against oxidative stress induced by salt stress.
6- Production of antioxidants and membrane protecting compounds help plants accumulated trehalose to withstand salt exposure. It is reasonable to believe that specific membrane lipid classes and sustainable transport proteins are constitutive or induced under saline conditions which might contribute to salt tolerance acquired by trehalose accumulation.
7- Exogenous application of validamycin A caused high accumulation of K+, N and P and lowering the content of Na+ in both cultivars under salt stress conditions.
Further investigations are needed to validate the effectiveness of this antibiotic manipulation under field conditions and to establish the effective doses based on different salt stress levels.
1- Salt stress-induced reduction in number of seeds, grain yield per plant, 1000-grain weight and filled-grain percentage for both rice cultivars was alleviated by exogenous application of validamycin A as pre-sowing seed treatment under saline field conditions
2- Exogenous application of validamycin A was found to be most effective at AgamiM5 cultivar as compared to the other cultivar Skha103.
At the molecular level, alterations in protein pattern of the treated rice tissues revealed that validamycin A treatments induced the appearance of new set of protein bands concomitant with the disappearance of others. Moreover increment in TPS gene expression of both rice cultivars tissues over those untreated tissues was recorded.
Other data
| Title | Physiological and Molecular Studies on Some Rice Cultivars | Other Titles | دراسات فسيولوجية و جزيئية على بعض أصناف الأرز. | Authors | Al Shymaa Zaky Al Mokadem | Issue Date | 2014 |
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