Genotyping safflower (Carthamus tinctorius) cultivars by DNA fingerprints

Summary Carthamus tinctorius (2n = 2x = 24) (family Asteraceae), commonly known as safflower, is widely cultivated in agricultural production systems of Asia, Europe, Australia and the Americas as a source of high-quality vegetable and industrial oil. India ranks first in the production of safflower oil. Fourteen cultivars, widely cultivated in various agro-climatic regions of India, have been fingerprinted by RAPD, ISSR, and AFLP markers utilizing 36, 21 primers, and 4 primer combinations, respectively. On an individual assay basis, AFLP has proven to be the best marker system as compared with the other two markers applied as assessed by high discriminating power (0.98), assay efficiency index (33.2), marker index (18.2), resolving power (40.62), and genotype index (0.856). Thirty-six RAPD and 21 SSR primers could differentiate a maximum of eight and four cultivars, respectively, whereas, two AFLP primer combinations could fingerprint all the 14 cultivars. To understand genetic relationships among these cultivars, Jaccard's similarity coefficient and UPGMA clustering algorithm were applied to the three marker data sets. Mean genetic similarities ranged from 0.689 (AFLP) to 0.952 (ISSR). Correlation coefficient comparisons between similarity matrices and co-phenetic matrices obtained with the three markers revealed that AFLP displayed no congruence vis-a-vis RAPD and ISSR data. However, strong correlation was observed between RAPD and ISSR marker systems. This paper reports the start of molecular biology programme targeting nuclear genome of safflower, a major world oilseed crop about whose genetics very little is known.     

First glimpse into the origin and spread of the Asian longhorned tick,Haemaphysalis longicornis,in the United States

Established populations of Asian longhorned ticks (ALT), Haemaphysalis longicornis, were first identified in the United States (US) in 2017 by sequencing the mitochondrial cytochrome c oxidase subunit I (cox1) ‘barcoding’ locus followed by morphological confirmation. Subsequent investigations detected ALT infestations in 12, mostly eastern, US states. To gain information on the origin and spread of US ALT, we (1) sequenced cox1 from ALT populations across 9 US states and (2) obtained cox1 sequences from potential source populations [China, Japan and Republic of Korea (ROK) as well as Australia, New Zealand and the Kingdom of Tonga (KOT)] both by sequencing and by downloading publicly available sequences in NCBI GenBank. Additionally, we conducted epidemiological investigations of properties near its initial detection locale in Hunterdon County, NJ, as well as a broader risk analysis for importation of ectoparasites into the area. In eastern Asian populations (China/Japan/ROK), we detected 35 cox1 haplotypes that neatly clustered into two clades with known bisexual versus parthenogenetic phenotypes. In Australia/New Zealand/KOT, we detected 10 cox1 haplotypes all falling within the parthenogenetic cluster. In the United States, we detected three differentially distributed cox1 haplotypes from the parthenogenetic cluster, supporting phenotypic evidence that US ALT are parthenogenetic. While none of the source populations examined had all three US cox1 haplotypes, a phylogeographic network analysis supports a northeast Asian source for the US populations. Within the United States, epidemiological investigations indicate ALT can be moved long distances by human transport of animals, such as horses and dogs, with smaller scale movements on wildlife. These results have relevant implications for efforts aimed at minimizing the spread of ALT in the United States and preventing additional exotic tick introductions.     

Diversity and relationships of multipurpose seabuckthorn (Hippophae L.) germplasm from the Indian Himalayas as assessed by AFLP and SAMPL markers

Seabuckthorn, a non-leguminous nodule bearing dioecious shrub, is a storehouse of neutraceutical, pharmaceutical and cosmetic usage. The 348 genotypes of Hippophae rhamnoides ssp. turkestanica, H. salicifolia and H. tibetana were collected from 194 locations at 46 major sites across?~1,500?km from north-east to north-west Himalayas, harboring one of the most harsh, highly variable climatic and ecological conditions and rugged rocky terrain in the range of 3,000–5,000?m altitude. Amplified fragment length polymorphism (AFLP) and selective amplification of microsatellite polymorphic loci (SAMPL) DNA markers were utilized to assess, the genetic diversity of total spectrum of Himalayan germplasm resources and interrelationships among Hippophae taxa. A total of 151, 50, and 41 AFLP loci were detected in Hippophae rhamnoides ssp. turkestanica, H. salicifolia and H. tibetana, respectively; of these, 92.6, 30.6 and 25.1% were polymorphic. For SAMPL, the number of markers and polymorphism for these species were, respectively, 77 (77.7%), 41 (41.4%), and 23 (23.2%). Nei’s genetic diversity and Shannon’s information index values revealed that populations of H. rhamnoides ssp. turkestanica in Sumur and Raling in Ladakh and Lahaul-Spiti, respectively, were the most diverse. In H. salicifolia and H. tibetana, the populations in Changu, and Takcha and Guling in Spiti valley, were the most diverse. In H. rhamnoides ssp. turkestanica, high level of interpopulation genetic diversity with the little intra-population diversity was accompanied by very low gene flow (Nm) range, estimated by AFLP (0.571–0.943) and SAMPL (0.321–0.726) markers. In H. rhamnoides ssp. turkestanica, the genotypes collected from Uttaranchal were found to be the least diverse. The exclusive characteristics of the nuclear genome in the Uttaranchal genotypes warrant new species rank more closer to H. salicifolia rather than to H. rhamnoides ssp. turkestanica. The present results also provide explicit evidence to suggest that both H. salicifolia and H. tibetana deserve species rank.     

Further heterogeneity of human alpha interferon mRNA species

Translationally active (in Xenopus oocytes) human alpha interferon (IFN) messenger RNA's (mRNA's) derived from Sendai virus--induced leukocyte cultures display a bimodal distribution of RNA lengths on electrophoresis through agarose-CH3HgOH gels. The major population (alpha s) consists of mRNA of length 0.7 to 1.4 kilobases, while the minor population (alpha L) consists of RNA of length 1.6 to 3.5 kilobases. Induction of human leukocytes in the presence of 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB; 100 micromolar) appears to inhibit the accumulation of IFN-alpha s and to enhance that of IFN-alpha L mRNA's (average length about 1.8 kilobases in preparations from DRB-treated cells). Interferons derived from the alpha s mRNA's represent the group of previously recognized alpha interferons while the alpha L interferons are distinguishable from this group by their lower heterospecific activity on bovine cells compared to human cells, their apparent slower mobility in sodium dodecyl sulfate--polyacrylamide gels, and their apparent heteroclitic response toward an antiserum to IFN-alpha.     

Composition and mineralogy of iron-manganese concretions from some soils of the Indo-Gangetic plain in Northwest India

Iron-manganese concretions from three reference soil series of the Indo-Gangetic plain of Punjab have been investigated for their composition, micromorphology and mineralogy. The data show that amounts of Fe, Mn, Co, Ni, Cu and Zn in the concretions are higher than in the surrounding soil matrix. The contents of Mn, Co and Ni increase and that of Fe decreases with the increasing size of concretions. Cu or Zn contents do not show any systematic differences with concretion size. A major portion of these elements is oxalate- and dithionite-extractable. A minor fraction is present in the silicate structure as well as in the pyrophosphate-extractable forms. The Fe and Mn compounds present in the concretions are x-ray amorphous. The concretions appear to have developed in situ.     

Genetic variation in wood specific gravity among half-sib families of chir pine (<Emphasis Type="Italic">Pinus roxburghii</Emphasis> sargent)

Chir pine (Pinus roxburghii) is an important tree species that grows all along the Himalayas. Wood specific gravity of chir pine before and after removal of extractives from wood increment cores was assessed from a 22-years-old progeny test in Himachal Pradesh, India. These values averaged 0.433 and 0.425, respectively. A large amount of genetic variation among the 58 half-sib families was found as indicated by range of values, additive coefficient of variation, variance estimates and narrow-sense heritability values for these traits. Moisture and extractive content averaged 86.259% of oven-dry weight and 2.003% of extractive-free oven-dry weight, respectively. Wood extractive content was highly variable and the family differences were highly heritable (h _f² = 0.5831). There was wide variability in moisture content, but a large portion of it was due to environmental or non-additive component of genetic variation. Heritability on family mean basis was found to be lower than that on individual tree basis for each trait. Estimated gain in specific gravity resulting from 30 to 50% family selection ranged from 0.0080 to 0.0127. Growth data and specific gravity were not significantly correlated implying that selection for higher growth rate would not necessarily result in reduction in wood specific gravity in chir pine.     

Vitellogenin genes in fish: differential expression on exposure to estradiol

Three types of vitellogenins (Vgs) namely vitellogenin A (VgA), vitellogenin B (VgB) and vitellogenin C (VgC) have been identified in fishes. The existence of VgA and VgB is reported in the Indian freshwater murrel Channa punctatus. Gene-specific primers were designed using available nucleotide sequences in National Centre for Biotechnology Information (NCBI), for amplification of VgA and VgB cDNA. Differential processing of Vgs is evident in many fishes. Adult male murrel expressed both the VgA and VgB genes when estradiol-17β (E₂) is injected in vivo and Vg levels in blood quantified by Enzyme linked immunosorbent assay (ELISA) showed a dose-related response in such treatments. Cultured hepatocytes on treatment with E₂, however, expressed only VgB as detected by RT-PCR, suggesting different regulatory mechanism for the VgA and VgB genes.     

Effect of domestic processing on total and extractable calcium and zinc content of Bathua ( Chenopodium album) and Fenugreek (Trigonella foenum graecum) leaves

Bathua (Chenopodium album) and fenugreek (Trigonella foenum graecum) stored in polyethylene bags and without packaging for 24 or 48 hours in a refrigerator at 5 or 30 ^°C in polyethylene bags. The fresh leaves were also dried (oven and sun); blanched (5, 10 or 15 min) and cooked in an open pan and a pressure cooker. The processed leaves were analyzed for total and extractable calcium and zinc content. The Ca and Zn content of these leaves varied from 970 to 2230 and 10.50 to 12.30 mg/100 g DM and the percentage HCl-extractability was 80.34 to 83.04 and 82.43 to 83.90, respectively. Non significant effects of drying and storage were observed on total Ca and Zn content and HCl-extractability while blanching and cooking resulted in significant improvement of HCl-extractability of these two minerals. Thus, cooking and blanching are good ways to improve the HCl-extractability of Ca and Zn.     

Biochemical changes in the liver of the Indian freshwater murrel, Channa punctatus (Bloch) during estradiol-induced vitellogenin synthesis

Estrogen administration in C. punctatus increases RNA: protein and RNA: DNA ratios and depletes glycogen in the liver. Increase in glucose-6-phosphatase activity accounts for glycogen depletion whereas high activity of pyruvate kinase suggests stimulation of the glycolytic pathway at the pyruvate step to generate ATP and to provide carbon skeleton for the vitellogenin molecule. Five-fold increase in the activity of NADP⁺-dependant malate dehydrogenase generates NADPH possibly for fatty acid synthesis. Additional energy requirement (NADPH) is met by stimulation of hexose monophosphate pathway as well as NADP⁺-dependant isocitrate dehydrogenase. Increase in the activity of NADPH-cytochrome c reductase in microsomal fraction suggests stimulation of electron transport chain. Increase in the activity of glutamate pyruvate transaminase can be directly related to the synthesis to specific amino acids needed for the formation of vitellogenin whereas reduction in glutamate oxaloacetate suggests extra hepatic source of amino acids. Reduction in the activity of glutamate dehydrogenase may be due to catabolism.