Search

Premature seed shattering in canola causes massive losses in yield by up to 50% in adverse climatic conditions. In the model plant Arabidopsis thaliana, which belongs to the same family as canola, the Brassicaceae, eight genes participate in a shattering cascade. Phylogenetic reconstruction, syntenic relationships, genomics loci, promoter sequences, and identification of transcription factor-binding sites (TFBSs) faced shattering cascade genes' analysis. Among these, three genes, SHATTERPROOF1, SHATTERPROOF2, and FRUITFUL (SHP1, SHP2, FUL), belonged to a MADS-box family implicated in fruit dehiscence zone and valve margin constitute a core regulatory module. But, in Brassica, the exact number of genes involved in shattering remained obscure. Grouping BnSHP1-N, BnSHP2-N, and BnFUL-N into their respective clades was according to phylogenetic reconstruction of core regulatory modules (SHP1, SHP2, and FUL) and from other species homologs. The eight shattering cascade genes showed no conservation, indicating their involvement in crushing through separate pathways. The increased number of homologs/paralogs in Brassica was due to occurrences of genome duplication or a triplication event during evolution. Exonization and intronization could be responsible for a variable number and size of the exons and introns in gene structures. Comparative genome synteny analysis of SHP1, SHP2, and FUL revealed correlation and evolutionary insights into gene region relationships in all Brassicaceae. Study results provided basic information on cloning, phylogenetic reconstruction, genomics loci, and identifying transcription factor-binding sites (TFBSs) of core regulatory module genes that might be helpful for developing shattering-resistant genome-edited plants to prevent future yield losses in canola. [ABSTRACT FROM AUTHOR]

The current study was based on Agrobacterium-mediated peanut genetic transformation and effective remediation of transgenic plants. The effect of Agrobacterium tumefaciens strain LBA4404 with a binary vector of PxCSG m-YFP 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene has been evaluated on two commercial varieties and on two sources of explants (Embryo and cotyledon), each containing a CaMV 35S promoter. The comparative regeneration and transformation efficiency analyses revealed that the embryo is a better target tissue than cotyledon, and early-ripe peanut genotypes with relatively small seeds (such as Bard-479) have proven to be comparatively responsive to transformation. Explant culture in media containing 3 mg L 1 indole-3-butyric acid and 0.1 mg L 1 napthalene acetic acid resulted in strong roots of virtually all transgenic plants. More than 87% of the transplanted plants were able to produce morphologically normal blooms and pods containing viable seeds. The phenotypic and genotypic monitoring of the EPSPS gene inheritance over two generations exhibited the desired 3:1 inheritance. Our findings suggest that Agrobacterium-mediated transformation is a feasible and valuable tool for peanut breeding and functional genomics research. [ABSTRACT FROM AUTHOR]

The extensive use of antibiotics in animal feed and development of antimicrobial resistance has led to the development of probiotics to improve the rate of livestock productivity. The present study was designed to isolate breed specific lactic acid bacterial (LAB) strains as animal probiotics. Sixty-eight gram-positive and catalase-negative strains were isolated, from the gastrointestinal tract and milk of Nilli Ravi buffaloes. For the selection of the buffalo probiotic, the viable capability of the isolated strains in a medium with different rumen conditions and antimicrobial activity were determined. Six strains (NMCC- PI, NMCC- PT, NMCC- P3n, NMCC- P7, NMCC- P8 and NMCC- F) showed a positive antibacterial as well as maximum cellular viability. These strains were identified as Enterococcus lactis (NMCC- PI) Pediococcus pentosaceus (NMCC- PT, MK014295), Enterococcus faecalis (NMCC-P3n) Lactobacillus fermentum (NMCC-P7), Pediococcus acidilactici (NMCC-P8) and Enterococcus ratti (NMCC-F) through 16 S rRNA gene sequencing. All strains fulfilled the basic criteria of animal probiotic properties however P. acidilactici (NMCC-P8) showed best result in terms of survivability at low pH (2.0), better antimicrobial activity and maximum enzymatic potential. Present results demonstrated that P. acidilactici may serve as a promising candidate probiotic for use in the buffalo dairy industry. This study was the unique to select LAB, with the potential to be used as probiotic from Nilli Ravi buffalo ruminal gut based on the ruminal-gut conditions. [ABSTRACT FROM AUTHOR]

This study was carried out to optimize in vitro regeneration system of groundnut through tissue culture technology that can provide good environment in terms of food security. The two varieties of groundnut (Golden and BARI 2001) were used and an effective in vitro regeneration system was established for selecting the best responsive variety to tissue culture using embryo as an explant sources. MS media having 5.5 mg/l BAP and 1.5 mg/l NAA was found to be best by producing the highest callus induction frequencies (86 & 78%) in Golden and BARI 2001, respectively. Similarly, the maximum multiple shoots/plant (9 & 6) with optimum length of shoots (4.8 & 4.1 cm) was obtained with the application of 4 mg/l BAP along with 1 mg/l NAA and 1.1 mg/l TDZ in Golden and BARI 2001, respectively. In case of roots initiation, the highest root frequency (90 & 76%) having optimum roots/shoots (8.3 & 6.2) and substantial root length (8.1 & 7.0 cm) was obtained in Golden followed by BARI 2001 variety on MS media having 1 mg/l phytohormone IBA. Results showed that callus and multiple shoot induction and in vitro rooting, varied with different hormonal combinations in the media and the genotype that might be due to genetic variations between the varieties. This study delivers a base line for efficient in vitro propagation and genetic transformation in elite groundnut genotypes for desirable characteristics. [ABSTRACT FROM AUTHOR]

A study was conducted to determine the status of gene combination for rust (Puccinia spp.) resistance in 100 indigenous wheat land races also yield related traits were evaluated at two different locations during the 2016-17 periods with Randomized Complete Block Design. Variations for the studied traits were conferred by Principal Component Analysis (PCA), PC1 with 80.1% variation for the Peshawar, and 93.8% for the Islamabad location. WMC419, XGWM120, GWM174, XGWM140, XGWM410, XGWM111, WMC773, XWMC170, XWMC405, XWMC348 and XWMC407 has no allelic variation because of monomorphic nature while XGWM35 (200&225 bp), GWM 148 (190&200 bp), Barc 86 (200&210 bp), BARC 114 (105&200 bp), CSLV34 (150&190 bp), PSP3000 (300&350 bp), XGWM493 (150&300 bp), XGWM153 (100&300 bp), XGWM44 with 120,285,500 & 700 bp, XBARC4 (90&200 bp), XGWM125 (150&190 bp) show variation in alleles. Analysis of Yellow rust (Yr5,7,10,15,18,19,26,29,34, P138 and Sr13,) and leaf rust (Lr19,34 and 49) revealed A and B resistance groups. Group A comprised of 36% of total accessions, while rest of the accessions (64%) were present in group B. The study revealed high level of diversity assessed through PCA and cluster analysis. Based on marker results, 23 wheat lines ranging 17-21 alleles in combination were identified. These lines have great potential to be used in rust resistance breeding programs. [ABSTRACT FROM AUTHOR]

Serriphidium quettense of the Asteraceae family is endemic to Balochistan, Pakistan. It has been used in traditional medicine for treating gastrointestinal diseases; while also having germicidal and antibacterial properties. However, no molecular and chemical characterization was demarcated earlier. Thus this study aimed to depict the genetic variability in two mountainous populations by molecular markers. Genetic variability assessment was carried out by polymorphic bands analysis to generate a dendrogram based on Sequential Agglomerative Hierarchical and Non-overlapping (SAHN) algorithm by Unweighted Pair Group Method of Arithmetic (UPGMA) means by using Numerical Taxonomy and Multivariate Analysis System (NTSYS) Pc version 2.01 for Cluster analysis. Chemical characterization was done by evaluating Total Phenolic Content (TPC) and flavonoids. A total of 68 amplified loci with 30 polymorphic were found with an average of 11.3 and 45% polymorphism. Cluster dendrogram based on similarity coefficient ranged from 0.285 to 0.785. Chemical vaiation was assesed by Agglomerative Hierarchical Clustering (AHC) based on dissimilarity matrix as indicated by two main clusters. Cluster 1 was represented by two sub clusters exhibited more chemical variation with in Zargoon population. Overall results obtained exhibited the diverse pattern of genetic and chemical variation. In conclusion, molecular marker profiling together with phytochemical variation of total phenolic and flavonoid content in all accessions of S. quettense and impact of ecological diversity on genetic and chemical variation can be used as base line study. Furthermore, implementation of conservation strategies of this endemic medicinal plant is highly recommended. [ABSTRACT FROM AUTHOR]

Genetic diversity is the base of biological polymorphism and also the basic part of biological diversity. Significant role is played by genetic diversity and paternity of germplasm in cotton breeding. Genetic diversity in representative sets of advanced cotton genotypes was studied at molecular level employing polymorphic simple sequence repeats. Of seventy-two advanced cotton genotypes, thirty microsatellites primer pairs were studied, 29 showed to be polymorphic. DNA markers amplified fragments of 100 to 300 bp in size. The polymorphism information contents ranged between 0.76 and 0.99. Cluster analysis clearly grouped the 72 genotypes into seven groups A, B, C, D, E, F, G based on Nei's coefficient similarity matrix. Group B was found to be the most genetically distinct group when compared to other groups. However, group B had most of the candidates of under observed genotypes. This study exposed two major clusters of cotton cultivars and recognized the cotton genotypes which were genetically different. These results can be used as reference and baseline information to other researchers/breeders. The resolution to the genetic associations of these cotton accessions could be improved by increasing the number of more mapped markers. [ABSTRACT FROM AUTHOR]

Drought stress hinders root and shoot growth, leaf area, biomass accumulation, plant fresh and dry weight signifying the severe losses of plant growth and development. During drought stress, different genes are activated that increase the drought tolerance efficiency of plants. In this study, DREB1A gene was transformed in wheat cv. Lasani-08 to improve drought tolerance efficiency after optimizing in planta Agrobacterium mediated transformation protocol. The results reveal that 3.0 mg/l 2, 4-D and 6 h soaking period were found to be the best combination for in planta transformation. The inoculation of seeds with Agrobacterium suspension before germination of apical meristem was proved to be the best time for obtaining more transformation efficiency (6.38%) as compared to seeds inoculation after germination of apical meristem. Molecular analyses of transgenic plants through PCR, Southern blotting and RT-PCR confirmed the presence, number of copies and expression profile of transgene in T1 generation. The drought tolerance efficiency was assessed in T2 transgenic plants on the basis of agronomical and biochemical analyses. These detailed analyses reveal that DREB1A gene has fostering effect on the expression of a large number of genes by means of enhanced germination frequency, germination rate index, shoot, root and coleoptiles lengths, relative water contents, proline contents and total chlorophyll contents of transgenic plants relative to control plants. In summary, our results propose that DREB1A is involved in improving drought stress tolerance of transgenic wheat by sustaining growth and yield of plants. [ABSTRACT FROM AUTHOR]

Walnut (Juglans regia L.) is exceptionally widespread nut tree in the world and grow best in Azad Jammu and Kashmir and northern areas of Pakistan is expected to have considerable inter and intra-species genetic variability due to adaptation to the various environmental conditions. We focused to evaluate the genetic characterization of sixty four walnut cultivars including their genetic relationship and distance collected from sixteen selected sites of Neelum valley Azad Jammu and Kashmir in Himalayan region of Pakistan. 35 decamer primers were used which based on discernable bands yielded markable amplification patterns. Cluster analysis and principal component analysis (PCA) separated the sixty four cultivars into six main groups whose alterations were related to their pedigree from each other. In our study we found 44% cultivars had marker alleles associated with the presence of WGA gene. 30% had at VAM gene. 18% had at JM gene. Six markers did not show polymorphism (were not validated). Frequencies of JM and VAM genes need to be improved in future walnut cultivars. Advanced walnut cultivars may routinely be investigated for the presence or absence of their special variety genes by the use of DNA markers. Kagazi akhrot need to be pyramided through MAS to extend the genetic base of future varieties. Moreover, fieldwork study revealed that local walnut germplasm are being eroded from some regions due to urbanization, agriculture, grazing, wood harvesting and natural calamities (earth quake and land sliding) that depicted genetic drift. The insightful difference among walnut accessions confirmed that they did not develop locally but were introduced from abroad germplasm whereas, close relationship showed that they are probably from same population with common ancestry. [ABSTRACT FROM AUTHOR]

Cotton is a cash crop of Pakistan and it plays a key role in the economy of the country for earning of foreign exchange. American cotton (Gossypium hirsutum L.) is the highest fibre producing specie of the world. The present study was conducted to check the comparison of morphological data, fibre quality traits and expression analysis of Cry1Ac gene in 14 cotton genotypes including hybrid (FI), F2 and F4 stages progeny in comparison with check variety MNH-886 at National Agricultural Research Centre, Islamabad during the year 2015 and 2016. The results of morphological traits revealed that field grown cotton genotypes showed variation in morphological characters. The highest plant height was observed 141.6 cm in HSP 7-4(2). Similarly, both maximum number of bolls per plant 115.60 and boll weight 5.92 g was reported in cotton hybrid NIGAB-4 respectively. The best internodes distance 2.32 cm was recorded in B-318-A genotype. There were significant differences observed in morphological traits of cotton except some genotypes. The results of fibre quality traits illustrated that optimum fibre fineness 3.1 µg/inch was reported in cotton hybrid NIGAB-4. While the best results of fibre strength 28.6 g/tex was measured in HSP 7-4(2) and hybrid NIGAB-3, length 33 mm in NIGAB-4, uniformity ratio 49.4 % in NIGAB-4 and fibre elongation 12.9 % was recorded in B1-37 genotype respectively. Meanwhile different Bt genes in 14 cotton genotypes (Cry1Ac, Cry2Ab and Cry1F genes) were checked through immunostrip assay and hence only Mon531 event for Cry1Ac gene of 346 bp was confirmed after immunostrip and PCR analysis. Quantification of Cry1Ac toxin was performed through sandwich ELISA at 80 and 120 days after sowing of leaf and boll tissues of 14 cotton genotypes. The Cry1Ac toxin level of these genotypes is in the range of 0.27 to 3.67µg/g based on fresh weight tissues. From the results, it has been concluded that cotton hybrids have performed well as compared to other tested cotton genotypes. [ABSTRACT FROM AUTHOR]

The yield of chickpea in Pakistan is unexpectedly low and unstable that may be due to biotic and abiotic stresses. To combat with these stresses, genetic make of chickpea should be improved incorporating important genes. For successful gene incorporation, an efficient tissue culture system may be established. Therefore, this study was conducted in two local chickpea genotypes namely Bittle-98 and Dasht-2000 for the establishment of an efficient tissue culture system. Among various concentrations of BAP, MS medium fortified with 3.0 mg/l BAP showed the highest multiple shoot formation (69.2 and 70.66%) in cvs Dasht-2000 and Bittle-98, respectively. Similarly, the highest elongation of regenerated shoots (58.33 and 91.66%) was recorded in Bittle-98 and Dasht-2000, respectively on MS media supplemented with 0.25 mg/l IAA. The highest rooting formation (60 and 80%) was recorded in Dasht-2000 and Bittle-98, respectively on MS media having 1.0 mg/l IBA. Half Embryo with single cotyledon was found to be the best explant for genetic transformation. During this study, hygromycin (75 mg/l), acetosyringone (100 μM) and cefotaxime sodium (500 mg/l) were found to be optimum for Agrobacterium-mediated transformation in chickpea. The putative transgenic plants were subjected to GUS and microscopic observations that confirmed the presence of uidA gene in both genotypes. The optimized protocols will provide a base line to improve transformation system and incorporate important genes in chickpea in future research programs. [ABSTRACT FROM AUTHOR]

Drought is a major limiting factor affecting productivity. Wheat is a major crop and staple food in Pakistan. Genetic linkage map construction based on linked DNA markers spanning whole wheat genome and subsequently QTL mapping for drought tolerance is a way forward to enhance breeder's ability for effective selection. An F8 population derived from the cross of OPATA x SH-349 has been used. An experiment was conducted at germination stage under controlled conditions. The drought was induced by15% PEG nutrient solution in acid washed sand medium. Microsatellite DNA markers data were used for linkage maps construction. Morphological data under drought and non-stressed conditions along with linkage maps data were used for QTL mapping. The results of QTL analysis using single marker analysis showed 14 SSR markers linked to QTLs for five traits in both drought and control condition. Using simple interval mapping and composite interval mapping 12 QTLs for different traits of interest were found. Implications of found QTLs and linked markers are discussed. [ABSTRACT FROM AUTHOR]

In vitro morphogenesis is greatly influenced by plant growth regulators (PGRs). The effect of different plant growth regulators was assessed on callus induction, in vitro shoot regeneration and hairy root formation in tomato. The maximum callus induction frequency was recorded culturing hypocotyls, while in vitro shoot regeneration frequency was significantly higher when leaf discs were used as explants. The highest callus induction frequency (67.48%) was recorded in cv. Rio Grande followed by Roma (62%) and Moneymaker (58.23%) on MS basal media along with 2.0 mg/l IAA and 2.5 mg/l BAP. Similarly, the highest in vitro shoot regeneration frequency (69.6 and 65.3%) was recorded in Rio Grande and Roma on MS media fortified with 0.1 mg/l IAA, 1.0 mg/l ZEA and 2.0 mg/l BAP, while in case of Moneymaker, the highest in vitro shoot regeneration frequency (67.3%) was obtained on MS media enriched with 0.1 mg/l IAA and 3.0 mg/l BAP. The highest hairy root formation frequency (100, 91.72 and 93.74%) was achieved in cvs. Rio Grande, Moneymaker and Roma, respectively on MS media supplemented with 0.1 mg/l IAA, 2.0 mg/l ZEA and 2.0 mg/l BAP. These optimized procedures would be applicable to other genotypes of tomato for the development of cultivars tolerant to abiotic stresses developed via Agrobacterium mediated transformation system. [ABSTRACT FROM AUTHOR]