A non-invasive method for phenotyping scab-tolerant apple plants using volatile organic compounds.

One of the most devastating diseases of apples is scab, caused by the fungus Venturia inaequalis. Most commercial apple varieties are susceptible to this disease; only a few are resistant. Breeding approaches are being used to develop better apple varieties that are resistant to scab. Volatile organic compounds (VOCs) contribute greatly to a plant's phenotype, and their emission profile largely depends on the genotype. In the non-destructive phenotyping of plants, VOCs can be used as biomarkers. In this study, we assessed non-destructively the scab tolerance potential of resistant (cv. 'Prima') and susceptible (cv. 'Oregon Spur') apple cultivars by comparing their major leaf VOC compositions and relative proportions. A comparison of the leaf VOC profiles of the two cultivars revealed 16 different VOCs, with cis-3-hexenyl acetate (3HA) emerging as a biomarker of cultivar differences. V. inaequalis growth was significantly inhibited in vitro by 3HA treatment. 3HA was significantly effective in reducing scab symptoms on V. inaequalis-inoculated leaves of 'Oregon Spur.' The resistant cultivar 'Prima' also exhibited higher lipoxygenase (LOX) activity and α-linolenic acid (ALA) levels, suggesting that V. inaequalis resistance is linked to LOX activity and 3HA biosynthesis. This study proposes 3HA as a potential biomarker for rapid non-destructive screening of scab-resistant apple germplasm of 'Prima' based on leaf VOCs.

Genetic diversity, population structure and marker-trait associations in Indian kale (Brassica oleracea L. gp. acephala) using cross-species microsatellite markers.

Kale is known for its exceptional nourishing and functional benefits to human body. However, it is an understudied species from genomic as well as agronomic aspects. It is important to characterize niche kale germplasms around the world to systematically conserve and utilize its genetic variability, especially for commercial traits in the interest of growers, consumers and industry. With this view, genomic and phenotypic characterizations of 62 Kashmiri kale accessions including popular landraces were done to estimate and partition genetic diversity, understand trait relationships, develop population structure and divulge marker-trait associations of economic significance. Sixty-six cross species microsatellite (SSR) markers within Brassica genus amplified 269 alleles in the germplasm. Their polymorphic information content (PIC) ranged from 0.00078 to 0.953 with an average of 0.407. The population structure analysis and neighbour joining tree clustering categorized the germplasm into three sub-populations. AMOVA revealed more within-population variance (67.73 %) than among-populations (32.27 %) variance. The principal component analysis (PCA) involving 24 agronomical traits revealed seven PCs (PC1 to PC7) having Eigen values more than 1, which explained a cumulative variation of 69.21 %. Association mapping with respect to these 24 agronomical traits using mixed linear model and general linear model revealed six overlapping significant marker-trait relationships with five being significant at probability value of 0.001/0.0001. The highly significant associations of two SSRs with economically important traits (siliqua length and seed weight) significantly correlated/related with leaf yield and seed yield were revealed for their possible utilization in marker assisted breeding for higher leaf and seed yields.

Flavor profiling and gene expression studies of indigenous aromatic rice variety (Mushk Budiji) grown at different altitudes of Highland Himalayan regions.

Mushk Budiji-an indigenous aromatic rice variety is usually grown at an altitude ranging from 5000 to 7000 ft above mean sea level in Highland Himalayas. This study was conducted to investigate the effects of altitude, soil nitrogen content and climatic conditions (temperature) of the selected locations on the flavor profile of Mushk Budiji using gas chromatography-mass spectroscopy (GC-MS) and electronic nose (E-nose). E-nose being rapid and non-destructive method was used to validate the results of volatile aromatic compounds obtained using GC-MS in Mushk Budiji. Around 35 aromatic compounds were identified in Mushk Budiji rice samples. Highest volatile peak area percentage (105.41%) was recorded for Mushk Budji grown at an altitude of 5216.53 ft. Highest E-nose score (2.52) was obtained at an altitude of 6299.21 ft. Over-expression of fatty acid degradation and linoleic acid metabolism genes was observed at higher altitudes, whereas lipid biosynthesis was negatively influenced by higher altitude. Fatty acid degradation and linoleic acid metabolism is responsible for the synthesis of volatile aromatic compounds in Mushk Budiji. This study will therefore be the path finder for investigating the intricate mechanism behind the role of altitude on aroma development in Mushk Budiji rice for future studies.

Methyl jasmonate improves resistance in scab-susceptible Red Delicious apple by altering ROS homeostasis and enhancing phenylpropanoid biosynthesis.

Apple (Malus domestica) is an economically important rosaceous fruit crop grown at temperate climate zones. Nevertheless, its production is severely affected by scab disease caused by the ascomycetous fungus Venturia inaequalis (VI). Methyl jasmonate (MeJA) is a stress induced plant hormone, shown to induce resistance against wide range of pathogens. The current study investigated the role of MeJA in promoting scab tolerance in susceptible apple varieties through exogenous application of optimized (100 µM) MeJA concentration, followed by VI infection. According to our analysis, applying MeJA exogenously onto leaf surfaces resulted in increased membrane stability and decreased malondialdehyde levels in Red Delicious, suggesting that MeJA is capable of protecting tissues against oxidative damage through its role in restoring membrane stability. In addition, the changes in the levels of key antioxidative enzymes and reactive oxygen species (ROS) showed that exogenous MeJA maintains ROS homeostasis as well. Higher phenylalanine ammonia-lyase activity and increased accumulation of phenylpropanoids in MeJA-treated VI-infected plants indicated the MeJA reprogrammed phenylpropanoid biosynthesis pathway for scab tolerance. Our study of scab tolerance in apples induced by MeJA provides new insights into its physiological and biochemical mechanisms.

Tissue and Time Optimization for Real-Time Detection of Apple Mosaic Virus and Apple Necrotic Mosaic Virus Associated with Mosaic Disease of Apple (Malus domestica).

Besides apple mosaic virus (ApMV), apple necrotic mosaic virus (ApNMV) has also been found to be associated with apple mosaic disease. Both viruses are unevenly distributed throughout the plant and their titer decreases variably with high temperatures, hence requiring proper tissue and time for early and real-time detection within plants. The present study was carried out to understand the distribution and titer of ApMV and ApNMV in apple trees from different plant parts (spatial) during different seasons (temporal) for the optimization of tissue and time for their timely detection. The Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) was carried out to detect and quantify both viruses in the various plant parts of apple trees during different seasons. Depending on the availability of tissue, both ApMV and ApNMV were detected in all the plant parts during the spring season using RT-PCR. During the summer, both viruses were detected only in seeds and fruits, whereas they were detected in leaves and pedicel during the autumn season. The RT-qPCR results showed that during the spring, the ApMV and ApNMV expression was higher in leaves, whereas in the summer and autumn, the titer was mostly detected in seeds and leaves, respectively. The leaves in the spring and autumn seasons and the seeds in the summer season can be used as detection tissues through RT-PCR for early and rapid detection of ApMV and ApNMV. This study was validated on 7 cultivars of apples infected with both viruses. This will help to accurately sample and index the planting material well ahead of time, which will aid in the production of virus-free, quality planting material.

Comparative study of TNF-α and vitamin D reveals a significant role of TNF-α in NSCLC in an ethnically conserved vitamin D deficient population.

Growing evidence has implicated tumor necrosis factor-alpha (TNF-α) as an important regulator of the tumor microenvironment. Moreover, various molecular epidemiological studies have proposed vitamin D deficiency to be a mediator of cancer progression. Here we comparatively analyzed the role of TNF-α and vitamin D in non-small cell lung cancer (NSCLC) in an ethnically conserved vitamin D deficient population. Confirmed NSCLC cases (n = 190) matchedfor age, gender, dwelling, and smoking against cancer-free healthy controls ((n = 200) were genotyped for TNF-α promoter polymorphisms (rs361525 and rs1800629) by PCR-RFLP. 48 NSCLC tumor and adjacent normal tissues were quantified for TNF-α mRNA expression by RT-qPCR. 48 NSCLC cases and 60 healthy controls were analyzed for TNF-α and vitamin D serum levels by ELISA and chemiluminescence respectively. Our study indicates thatrs361525 and rs1800629 bear a significant risk towards NSCLC. Both mutant genotype and mutant allele of rs361525 elicit a likelihood of NSCLC reflected by their odds ratio (OR) of 3.16 and 1.81 respectively. In case of rs1800629, the heterogeneous genotype (GA) showed two fold higher risk for NSCLC (OR-2.07, P = 0.006), which could be attributed to the presence of the mutant allele as reflected by overall frequency of mutant A allele vs wild G allele (OR-1.92, P = 0.01). A combined effect of genotypes for rs361525 and rs1800629 revealed a 3.7 fold higher risk towards NSCLC for the presence of heterozygous genotype at both loci. Our preliminary expression results showed significant increase of TNF-α mRNA expression in tumor tissues of NSCLC as compared to adjacent normal tissues [cases- 8.56 ± 3.90vs controls-4.88 ± 2.96, P < 0.0001)] which was further affirmed by extrapolation of TNF-α expression in serum (Cases- 55.75 ± 22.50vs controls- 21.46 ± 27.75, P < 0.0001). Multivariate regression analyses revealed TNF-α mRNA expression to be significantly associated with NSCLC cases less than 50 years of age (P < 0.05). In comparison to the putative role of TNF-α in NSCLC as suggested by the results observed, vitamin D showed no significance towards any of the analyzed parameters or with the risk of NSCLC. This study suggests that TNF-α could be a potential mediator of NSCLC which bears important clinical implications and could be an important therapeutic marker in NSCLC.

SSR markers in revealing extent of genetic diversity and phylogenetic relationships among chickpea core collection accessions for Western Himalayas.

BACKGROUND: The exploration of genetic diversity is the key source of germplasm conservation and potential to broaden its genetic base. The globally growing demand for chickpea suggests superior/climate-resilient varieties, which in turn necessitates the germplasm characterization to unravel underlying genetic variation. METHODOLOGY AND RESULTS: A chickpea core collection comprising of diverse 192 accessions which include cultivated Cicer arietinum, and wild C. reticulatum, C. echinospermum, and C. microphyllum species were investigated to analyze their genetic diversity and relationship, by assaying 33 unlinked simple sequence repeat (SSR) markers. The results amplified a total of 323 alleles (Na), ranging from 2 to 8 with an average of 4.25 alleles per locus. Expected heterozygosity (He) differed from 0.46 to 0.86 with an average of 0.68. Polymorphic information content (PIC) ranged from 0.73 to 0.98 with an average of 0.89. Analysis of molecular variance (AMOVA) showed that most of the variation was among individuals (87%). Cluster analysis resulted in the formation of four distinct clusters. Cluster I represented all cultivated and clusters II, III, and IV comprised a heterogeneous group of cultivated and wild chickpea accessions. CONCLUSION: We report considerable diversity and greater resolving power of SSR markers for assessing variability and interrelationship among the chickpea accessions. The chickpea core is expected to be an efficient resource for breeders for broadening the chickpea genetic base and could be useful for selective breeding of desirable traits and in the identification of target genes for genomics-assisted breeding.

Elucidating Genetic Diversity in Apricot (Prunus armeniaca L.) Cultivated in the North-Western Himalayan Provinces of India Using SSR Markers.

Apricot (Prunus armeniaca L.) is an important temperate fruit crop worldwide. The availability of wild apricot germplasm and its characterization through genomic studies can guide us towards its conservation, increasing productivity and nutritional composition. Therefore, in this study, we carried out the genomic characterization of 50 phenotypically variable accessions by using SSR markers in the erstwhile States of Jammu and Kashmir to reveal genetic variability among accessions and their genetic associations. The genetic parameter results revealed that the number of alleles per locus (Na) ranged from 1 to 6 with a mean Na value of 3.89 and the mean effective number of alleles (Ne) per locus 1.882 with a range of 1.22 to 2. Similarly, the polymorphic information content (PIC) values ranged from 0.464 to 0.104. The observed heterozygosity (Ho) (0.547) was found to have higher than expected heterozygosity (He) (0.453) with average heterozygosity of 0.4483. The dendrogram clustered genotypes into three main clades based on their pedigree. The population structure revealed IV sub-populations with all admixtures except the III sub-population, which was mainly formed of exotic cultivars. The average expected heterozygosity (He) and population differentiation within four sub-populations was 1.78 and 0.04, respectively, and explained 95.0% of the total genetic variance in the population. The results revealed that the SSR marker studies could easily decrypt the genetic variability present within the germplasm, which may form the base for the establishment of good gene banks by reducing redundancy of germplasm, selection of parents for any breeding program.

The influence of COVID-19 related psychological and demographic variables on the effectiveness of e-learning among health care students in the southern region of Saudi Arabia.

BACKGROUND: E-learning is a modern and flexible mode of education and is being used as an alternative to conventional mode of education during the ongoing COVID-19 pandemic. However evidence suggests that effectiveness of e-learning is influenced by many prevailing factors. METHODS: A cross sectional study aimed to evaluate health care students' perception towards implementation of e-learning was conducted for a period of 4 months, from April 2020 to July 2020. Research instrument consisted of a self-designed, qualitative questionnaire with three domains was validated using field pretest method and administered among health care students of King Khalid University using social media platforms. Chi square test was used to estimate the effect of e-learning on various domains whereas linear regression analysis was used to find the association between subjects' characteristics to overall domain scores. P value < 0.05 was considered significant. RESULTS: Among 254 respondents 59.8% were males and 96.5% respondents were staying with their families.10.2% reported medically imposed quarantine for one or more family members. 59.8% reported that the online classes were commenced too early. 63.4% of the respondents reported that they had no previous experience. Poor network connectivity (32.3%) and unawareness about online platforms (29.9%), poor audio/video qualities (26.3%) were found as major barriers. Psychological stress, discipline of study, living status and quarantine history had a remarkable impact on the effectiveness of online education. CONCLUSION: Health care students are still cynical and are yet to embrace e-learning fully. Psychological distress, technical issues in association with accessibility, inexperience and unpreparedness were found to be main barriers that limited student acceptance of e-learning.

Plummeting anthropogenic environmental degradation by amending nutrient-N input method in saffron growing soils of north-west Himalayas.

Nitrous-oxide emission and nitrate addition from agriculture to earth's environment are two main agriculture related anthropogenic causes of environmental degradation that needs greater attention. For addressing the aforesaid issue, new techniques/practices need to be developed and implemented. The present investigation, which was focused on this issue, resulted in developing a new mode of nitrogen (N) placement, i.e. 'mid rib placement upper to corms in two splits (MRPU-2S)', that could reduce nitrous oxide N emission by around 70.11% and, nitrate N leaching and runoff by around 68.26 and 67.09%, respectively, over conventional method, in saffron growing soils of northwest Himalayas. Besides plummeting environmental degradation, MRPU-2S further resulted in enhancing saffron yield by 33.33% over conventional method. The findings of the present investigation were used to develop new empirical models for predicting saffron yield, nitrate N leaching and nitrous-oxide N emission. The threshold limits of nitrate N leaching and nitrous oxide N emission have also been reported exclusively in the present study.

Comparative evaluation of biphasic insulin with metformin and triple oral hypoglycemic agents (OHA) in type 2 diabetes patients.

INTRODUCTION: The prevalence of secondary failure to oral hypoglycemic agents among type 2 diabetes mellitus (T2DM) patients ranges from 30 to 60%. The alternative approaches to overcome this issue are either switching to triple oral hypoglycemic agents (OHA) or intensifying the regimen by adding insulin. OBJECTIVE: To compare the glycemic control achieved with biphasic insulin plus metformin and triple OHA in T2DM patients who were not adequately controlled with two OHA regimen. METHODS: A qualitative prospective study was conducted at Asir diabetes center, Abha, KSA. Poorly controlled T2DM patients with two OHA for at least 1 year with glycated hemoglobin (HbA1c) >7.0% were included. Subjects were divided into group I (a third OHA was added to the existing two OHA regimen) and group II (switched over to Biphasic insulin and metformin). At baseline and 3-month intervals, level of HbA1C, Fasting Plasma Glucose (FPG), Postprandial Plasma Glucose (PPG), Blood Pressure (BP), lipid profile and hypoglycemic episodes were obtained and evaluated for one year. RESULTS: 41.1% of patients were in group I and 58.9% were in group II. At the end of the study, there was a significant reduction in HbA1c in group II subjects comparing to group I (8.18 ± 1.32 vs 8.79 ± 1.81, p = 0.0238). FPG and PPG were improved also in group II. The mean body weight increased from baseline in group II is +4.48 kg and decreased from baseline in group I (-0.46 kg). 11.3% from group I and 23.7% from group II reported hypoglycaemic incidences. CONCLUSION: Biphasic insulin and metformin regimen could be an appropriate therapeutic option for achieving good glycemic control compared with triple OHA in patients with two OHA failure.

Cinnamate-CoA ligase is involved in biosynthesis of benzoate-derived biphenyl phytoalexin in Malus × domestica 'Golden Delicious' cell cultures.

Apple (Malus sp.) and other genera belonging to the sub-tribe Malinae of the Rosaceae family produce unique benzoic acid-derived biphenyl phytoalexins. Cell cultures of Malus domestica cv. 'Golden Delicious' accumulate two biphenyl phytoalexins, aucuparin and noraucuparin, in response to the addition of a Venturia inaequalis elicitor (VIE). In this study, we isolated and expressed a cinnamate-CoA ligase (CNL)-encoding sequence from VIE-treated cell cultures of cv. 'Golden Delicious' (M. domestica CNL; MdCNL). MdCNL catalyses the conversion of cinnamic acid into cinnamoyl-CoA, which is subsequently converted to biphenyls. MdCNL failed to accept benzoic acid as a substrate. When scab-resistant (cv. 'Shireen') and moderately scab-susceptible (cv. 'Golden Delicious') apple cultivars were challenged with the V. inaequalis scab fungus, an increase in MdCNL transcript levels was observed in internodal regions. The increase in MdCNL transcript levels could conceivably correlate with the pattern of accumulation of biphenyls. The C-terminal signal in the MdCNL protein directed its N-terminal reporter fusion to peroxisomes in Nicotiana benthamiana leaves. Thus, this report records the cloning and characterisation of a cinnamoyl-CoA-forming enzyme from apple via a series of in vivo and in vitro studies. Defining the key step of phytoalexin formation in apple provides a biotechnological tool for engineering elite cultivars with improved resistance.

Variability in Catechin and Rutin Contents and Their Antioxidant Potential in Diverse Apple Genotypes.

Catechins and rutin are among the main metabolites found in apple fruit. Sixty apple genotypes, harvested in 2016 and 2017, were analyzed for their phenolic content and antioxidant activity. The HPLC analysis showed that the catechin concentration ranged from 109.98 to 5290.47 µg/g, and the rutin concentration ranged from 12.136 to 483.89 µg/g of apple fruit. The level of DPPH activity ranged from 9.04% to 77.57%, and almost half of the 15 genotypes showed below 30⁻40% DPPH activity. The apple genotypes 'Lal Ambri', 'Green Sleeves', and 'Mallus floribunda' showed the highest DPPH activity of between 70% and 80%, while 'Schlomit', 'Luxtons Fortune', 'Mayaan', 'Ananas Retrine', and 'Chaubatia ambrose' showed the lowest ferric reducing antioxidant power (FRAP) activity (0.02⁻0.09%). Statistical analysis showed a correlation between DPPH activity and catechin content (r = 0.7348) and rutin content (r = 0.1442). Regarding antioxidant activity, fractionated samples of apple genotypes revealed significant activity comparable to that of ascorbic acid. There was also a consistent trend for FRAP activity among all apple genotypes and a significant positive correlation between FRAP activity and rutin content (r = 0.244). Thus, this study reveals a significant variation in antioxidant potential among apple genotypes. This data could be useful for the development of new apple varieties with added phytochemicals by conventional and modern breeders.

Molecular cloning and functional analysis of a biphenyl phytoalexin-specific O-methyltransferase from apple cell suspension cultures.

MAIN CONCLUSION: This manuscript describes the cloning and functional characterization of a biphenyl phytoalexin biosynthetic gene, 3,5-dihydroxybiphenyl O-methyltransferase from elicitor-treated cell cultures of scab resistant apple cultivar 'Florina'. Apples belong to the subtribe Malinae of the Rosaceae family. Biphenyls and dibenzofurans are the specialized phytoalexins of Malinae, of which aucuparin is the most widely distributed biphenyl. The precursor of aucuparin, 3,5-dihydroxybiphenyl, is a benzoate-derived polyketide, which is formed by the sequential condensation of three molecules of malonyl-CoA and one molecule of benzoyl-CoA in a reaction catalyzed by biphenyl synthase (BIS). This 3,5-dihydroxybiphenyl then undergoes sequential 5-O-methylation, 4-hydroxylation, and finally 3-O-methylation to form aucuparin. A cDNA encoding O-methyltransferase (OMT) was isolated and functionally characterized from the cell cultures of scab-resistant apple cultivar 'Florina' (Malus domestica cultivar 'Florina'; MdOMT) after treatment with elicitor prepared from the apple scab causing fungus Venturia inaequalis. MdOMT catalyzed the regiospecific O-methylation of 3,5-dihydroxybiphenyl at the 5-position to form 3-hydroxy-5-methoxybiphenyl. The enzyme showed absolute substrate preference for 3,5-dihydroxybiphenyl. The elicitor-treated apple cell cultures showed transient increases in the MdOMT (GenBank ID MF740747) and MdBIS3 (GenBank ID JQ390523) transcript levels followed by the accumulation of biphenyls (aucuparin and noraucuparin) and dibenzofuran (eriobofuran) phytoalexins. MdOMT fused with N- and C-terminal yellow fluorescent protein showed cytoplasmic localization in the epidermis of Nicotiana benthamiana leaves. In scab inoculated greenhouse-grown 'Florina' plants, the expression of MdOMT was transiently induced in the stem followed by the accumulation of biphenyl phytoalexins.

Comparative metabolomics of scab-resistant and susceptible apple cell cultures in response to scab fungus elicitor treatment.

Apple scab disease caused by the fungus Venturia inaequalis is a devastating disease that seriously affects quality and yield of apples. In order to understand the mechanisms involved in scab resistance, we performed gas chromatography-mass spectrometry based metabolomics analysis of the cell culture of scab resistant cultivar 'Florina' and scab susceptible cultivar 'Vista Bella' both prior -to and -following treatment with V. inaequalis elicitor (VIE). A total 21 metabolites were identified to be altered significantly in 'Florina' cell cultures upon VIE-treatment. Among 21 metabolites, formation of three new specialized metabolites aucuparin, noraucuparin and eriobofuran were observed only in resistant cultivar 'Florina' after the elicitor treatment. The score plots of principal component analysis (PCA) exhibited clear discrimination between untreated and VIE-treated samples. The alteration in metabolite levels correlated well with the changes in the transcript levels of selected secondary metabolite biosynthesis genes. Aucuparin, noraucuparin and eriobofuran isolated from the 'Florina' cultures showed significant inhibitory effect on the conidial germination of V. inaequalis. The results expand our understanding of the metabolic basis of scab-resistance in apple and therefore are of interest in apple breeding programs to fortify scab resistance potential of commercially grown apple cultivars.

Effect of environmental variables on phytonutrients of Origanum vulgare L. in the sub-humid region of the northwestern Himalayas.

Ecological and soil physiochemical parameters impact the crop quality and development. In spite of the huge commercial prospective, the phytonutrient and chemometric profiles of Himalayan oregano (Origanum vulgare L.) have not been evaluated, and their relationships with ecological parameters are still lacking. The objective of this research study was to evaluate the disparity in the phytonutrient profiles of different ecotypes of O. vulgare in wild and cultivated populations and determine whether such variation was related to the diverse climatic and edaphic conditions prevailing in the northwestern Himalayas. Micrometeorological, atomic absorption spectroscopy for micro-elemental analysis was determined for soil. HPLC was used to determine the disparity in phytonutrient (quercetin, betacarotene, ascorbic acid, and catechin) and phytochemical (arbutin) levels. Cultivated populations had lower phytonutrient levels than wild populations. The habitat exhibiting pH values ranging from 6 to 7 elevated organic carbon (2.42%), nitrogen (97.41 kg ha-1), and manganese (10-12 µg g-1) and zinc contents (0.39-0.50%) show luxirant growth of Origanum vulgarel. The phytonutrient (quercetin, betacarotene, ascorbic acid, arbutin, and catechin) levels had a direct relationship with UV-B flux (r2 = 0.82) and potassium (r2 = 0.97). Wild accessions predominantly contained catechin and ascorbic acid, with maximum values of 163.8 and 46.88 µg g-1, respectively, while the cultivated accessions had the highest level of arbutin (53.42 µg g-1). Maximum variation was observed in quercetin (114.61%) followed by ß-carotene (87.53%). Cultivated accessions had less quercetin (0.04-1.25 µg g-1) than wild accessions (1.25-2.87 µg g-1). Wild accessions had higher phytonutrient values for catechin, ß-carotene, and ascorbic acid while cultivated accessions had maximum values for arbutin. The correlation of environmental variables with phytonutrient levels paves the way for metabolomic-guided enhancement of agricultural practices for better herb quality.

Bioefficacy potential of different genotypes of walnut Juglans regia L.

The purpose of investigation was to assess the phytochemical and nutraceutical of walnut in leaf extracts through diverse quantitative and qualitative phytochemical tests followed by array of assays. The screening of 50 elite walnut genotypes which exhibited wide range of discrepancy in terms of phytochemicals as well as their anti-oxidant potential was done. Walnut genotypes displayed maximum divergence in quercetin content (2.86-5.78 mg/100 g) as represented by cluster analysis. The phenolic rich genotypes exhibiting total phenols (37.61-46.47 mg/g GAE) having higher DPPH potential (IP of 32.82-73.50) where as genotypes that accumulate flavonoids/flavanols (5.52-28.48 mg/g QE and 4.11-21.76 mg/g QE showed immense FRAP activity (418.92-1067.94 µM Fe2+/g FW). There was positive correlation between the phenolics content and anti-oxidant potential. The results showed oil content of 50.1-85.08% and kernel percentage 25.21-81.92% of all walnut genotypes. To evaluate the anti-proliferative potential of walnut genotypes, Trypan blue exclusion test, MTT assay and Griess assay was used. Each assay was repeated with different positive controls against a panel of human cancer cell lines viz THP-1, U2OS, IMR-32 and HBL-100 and then compared with the walnut extracts for their efficiency in anti-proliferative activity. The SPS 1 walnut extract at concentration of 500 µg/ml exhibited 10% cell viability and with 1000 µg/ml walnut extract there was consequent decline towards (6.25%) viability. The results indicated that walnut leaf constitutes an excellent source of effective natural antioxidants and chemo-preventive agents that can act as anti cancer agents.

Gene/QTL discovery for Anthracnose in common bean (Phaseolus vulgaris L.) from North-western Himalayas.

Common bean (Phaseolus vulgaris L.) is one of the most important grain legume crops in the world. The beans grown in north-western Himalayas possess huge diversity for seed color, shape and size but are mostly susceptible to Anthracnose disease caused by seed born fungus Colletotrichum lindemuthianum. Dozens of QTLs/genes have been already identified for this disease in common bean world-wide. However, this is the first report of gene/QTL discovery for Anthracnose using bean germplasm from north-western Himalayas of state Jammu & Kashmir, India. A core set of 96 bean lines comprising 54 indigenous local landraces from 11 hot-spots and 42 exotic lines from 10 different countries were phenotyped at two locations (SKUAST-Jammu and Bhaderwah, Jammu) for Anthracnose resistance. The core set was also genotyped with genome-wide (91) random and trait linked SSR markers. The study of marker-trait associations (MTAs) led to the identification of 10 QTLs/genes for Anthracnose resistance. Among the 10 QTLs/genes identified, two MTAs are stable (BM45 & BM211), two MTAs (PVctt1 & BM211) are major explaining more than 20% phenotypic variation for Anthracnose and one MTA (BM211) is both stable and major. Six (06) genomic regions are reported for the first time, while as four (04) genomic regions validated the already known QTL/gene regions/clusters for Anthracnose. The major, stable and validated markers reported during the present study associated with Anthracnose resistance will prove useful in common bean molecular breeding programs aimed at enhancing Anthracnose resistance of local bean landraces grown in north-western Himalayas of state Jammu and Kashmir.

In vitro development of microcorms and stigma like structures in saffron (Crocus sativus L.).

Saffron is an important spice derived from the stigmas of Crocus sativus, a species belonging to the family Iridaceae. Due to its triploid nature it is sterile and is not able to set seeds, so it is propagated only by corms. The natural propagation rate of most geophytes including saffron is relatively low. An in vitro multiplication technique like micropropagation has been used for the propagation of saffron. In the present study, various explants were cultured on different nutrient media supplemented with various concentrations of plant growth regulators to standardize the best media combination for obtaining optimum response with respect to corm production and development of Stigma Like Structures (SLS). Highest response (60 %) was observed with half ovaries on G-5 media supplemented with 27 µM NAA and 44.4 µM BA followed by 55 % on LS media with 27 µM NAA and 44.4 µM BA. Maximum size (1.3 g) of microcorms were obtained from apical buds on the LS media supplemented with 21.6 µM NAA and 22.2 µM. Stigma Like Structures were developed from half ovary explants both directly and indirectly. Maximum number (120 indirectly and 20 directly) and size (5.2 cm) of SLS were obtained in G-5 medium supplemented with 27 µM NAA and 44.4 µM BA followed by 100 indirectly and 20 directly and 4.5 cm long on LS medium supplemented with 27 µM NAA and 44.4 µM BA.