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Increased soil salinity in the recent years has adversely affected the productivity of mango globally. Extending the cultivation of mango in salt affected regions warrants the use of salinity tolerant/resistant rootstocks. However, the lack of sufficient genomic and transcriptomic information impedes comprehensive research at the molecular level. We employed RNA sequencing-based transcriptome analysis to gain insight into molecular response to salt stress by using two polyembryonic mango genotypes with contrasting response to salt stress viz., salt tolerant Turpentine and salt susceptible Mylepelian. RNA sequencing by Novaseq6000 resulted in a total of 2795088, 17535948, 7813704 and 5544894 clean reads in Mylepelian treated (MT), Mylepelian control (MC), Turpentine treated (TT) and Turpentine control (TC) respectively. In total, 7169 unigenes annotated against all the five public databases, including NR, NT, PFAM, KOG, Swissport, KEGG and GO. Further, maximum number of differentially expressed genes were found between MT and MC (2106) followed by MT vs TT (1158) and TT and TC (587). The differentially expressed genes under different treatment levels included transcription factors (bZIP, NAC, bHLH), genes involved in Calcium-dependent protein kinases (CDPKs), ABA biosynthesis, Photosynthesis etc. Expression of few of these genes was experimentally validated through quantitative real-time PCR (qRT-PCR) and contrasting expression pattern of Auxin Response Factor 2 (ARF2), Late Embryogenesis Abundant (LEA) and CDPK genes were observed between Turpentine and Mylepelian. The results of this study will be useful in understanding the molecular mechanism underlying salt tolerance in mango which can serve as valuable baseline information to generate new targets in mango breeding for salt tolerance.

IntroductionPutative mutants were generated through gamma irradiation in the polyembryonic mango genotype Nekkare. The putative mutant progenies along with control seedlings and mother plants were evaluated by comparing the compositions and relative proportions of their major volatile compounds.MethodsVolatile profiling was done using headspace-solid phase micro-extraction (HS SPME) method coupled with gas chromatography–mass spectrometry (GC-MS MS). Furthermore, characterisation of putative mutants and control seedlings was carried out using simple sequence repeat (SSR) markers to ascertain the genetic diversity present in the samples under study.ResultsMonoterpenes were the most abundant volatile compound in all the studied samples (ranging from 34.76% to 91.41%) out of which I-Phellandrene and cis-Ocimene formed the major fraction in mother plants (20.45%–21.86% and 16.17%–21.27%, respectively) and control seedlings (23.32%–24.95% and 18.95%–20.81%, respectively), while beta-Phellandrene was dominant in the selected putative mutant samples (2.34%–29.53%). Among sesquiterpenes, trans-Caryophyllene was detected only in the putative mutant samples (0.10%–30.18%). Grouping together of mother plants and control seedlings was seen in the cluster analysis, while the putative mutants grouped apart from them suggesting genetic diversity. Genetic distance between the mother plants and control seedlings ranged from 0.97 to 2.73, while between putative mutants, control seedlings, and mother plants, it ranged from 6.54 to 9.82. SSR-based characterisation of putative mutant seedlings showed that mutation caused variability in the treated population. This was evident from the high allelic richness ranging from 4 to 12 with a mean of 7 and a higher mean Shannon’s Information Index (1.50) of the putative mutant population.DiscussionThe study demonstrates that volatile profiling and molecular characterisation using SSR markers could be used as a tool to detect variation in a mutated population. In addition, volatile profiling can be used to validate putative mutants in polyembryonic mango genotypes where the seedlings of nucellar origin are similar to mother plants.

Urban environment and air quality are changing primarily due to land use land cover (LULC) changes, economic activity, and urbanization. Air pollution has been increasingly acknowledged as a major issue for cities due to its extensive effects on health and well-being. As the second most populous city in the country, Lahore faces alarming levels of air pollutants, which induced this study to focus on the pervasive issue of air pollution in Lahore. For this, the study collected air pollutants data from the Environmental Protection Department of Punjab and analyzed them using the ARIMA model. In the research results, both the observed data and predictive models uncovered concerning trends in pollutant concentrations, ultimately portraying a concerning picture for air quality management. Carbon monoxide (CO) levels show a consistent rise, surpassing Pakistan’s environmental standards by 2025. Similarly, nitrogen dioxide (NO 2 ) concentrations escalate, exceeding prescribed standards. Ground-level ozone (O 3 ) also demonstrates a substantial increase, surpassing standards by 2025. Both PM 2.5 and PM 10 exhibit marked upward trends, projected to exceed recommended limits, particularly PM 10 throughout the study year. The Air Quality Index exhibits an observable upward trend, fluctuating between 70 and 442 from 2015 to 2020. Similarly, a positive correlation was found between population growth and land use conversion into residential areas. Projections suggest a continuous increase, potentially hitting a severe level of 500 during winter by 2025. These findings point to an impending air pollution crisis, demanding urgent action to address the hazardous situation in the city. The study recommends that urban air pollution should be reduced, and the negative health effects of air pollution should be minimized using vegetation barriers, screens, and greening initiatives. Strict regulations and monitoring initiatives need to be put in place in big cities to monitor pollution and vegetation.

The effect of salt stress was evaluated on putative mutant seedlings of three polyembryonic mango genotypes viz., Bappakkai, Nekkare, and Kurukkan. Imposition of salinity stress resulted in a decrease in chlorophyll content, relative water content, and gas exchange parameters while enhancing the levels of stress markers like Na + /K + ratio, total phenols, and proline. In total, thirteen phenolic acid compounds were identified including eight hydroxybenzoic acids and five hydroxycinnamic acids wherein hydroxybenzoic acid (majorly gallic acid) comprised more than 99% of total phenolic acids. In all the three genotypes, the concentration of protocatechuic acid, 2, 4-Dihydroxy benzoic acid, gallic acid, chlorogenic acid, and t-cinnamic acid increased with increasing level of salt stress indicating their potential role in mango salt tolerance. Bappakkai recorded higher K + , highest fold increase in proline content (+ 7.27 fold), highest percent increase in chlorogenic acid (+ 510%), protocatechuic acid (+ 750%), and ferulic acid along with lower Na + /K + ratio and lower reduction in the levels of caffeic and sinapic acid at higher level of salt stress suggesting that putative mutants of Bappakkai were better at tolerating salt stress as compared to the other two genotypes. Exogenous application of ferulic acid (FA) to Nekkare putative mutants increased the activity of enzymatic antioxidants, superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). Further, quantitative real-time PCR analysis also revealed higher expression levels of SOD, CAT, and APX in FA-treated Nekkare mutants under salinity stress. The results of this study besides identifying distinct phenolic acid signatures in response to stresses like gamma irradiation and salinity, also confirm the potential of physical mutagenesis in breeding salt-tolerant rootstocks in mango.

The genetic diversity of 25 pomegranate genotypes, including widespread genotypes, mutants, clonal selections and wild forms, was assessed using 23 pomegranate microsatellite markers in this study. A total of 117 alleles were found, with 5.09 ± 0.47 alleles per locus on average. The number of alleles per marker (Na) ranged from 2 to 10, demonstrating that these microsatellites are highly informative. The predicted and actual heterozygosity mean values were 0.55 and 0.34, respectively. The polymorphic information content of selected SSR loci ranged from 0.08 to 0.80, with an average of 0.51, showing its usefulness in distinguishing the genotypes under study. Based on the PIC value, the locus PGKVR 153 was discovered to have the highest polymorphism levels (0.80). The 25 genotypes were divided into two main clusters and multiple sub and sub-sub clusters within the main clusters using the un-weighted pair-group method with arithmetic average (UPGMA) approach. Principle coordinate analysis (PCoA) clustering of genotypes was also consistent with UPGMA clustering. The findings confirm that the genotypes have a high genetic diversity and a broad genetic background, indicating that their inclusion into future crop breeding programmes would be successful.

Mango, one of the most important fruit crops in the world, has both monoembryonic and polyembryonic varieties. The nucellar seedlings from polyembryonic varieties are exactly the same as the mother plant, while zygotic seedlings vary due to fertilization of a zygote by the pollen parent. However, there are no reliable methods that include the molecular tools for the identification of nucellar seedlings. Therefore, leaf volatiles of polyembryonic and monoembryonic varieties were used for comparison with that of their respective seedlings. The leaf volatile constituents were analyzed using gas chromatography–mass spectrometry (GC-MS) by headspace solid-phase microextraction technique (HS-SPME). Volatile constituents of the last emerging seedling from a stone were different from those of the respective mother plant, while those of the remaining seedlings were exactly the same as each other and as the respective mother plant in polyembryonic genotypes, namely ‘Chengavarkai’, ‘Turpentine’ and ‘Gomavu’. In the case of monoembryonic cv. ‘Totapuri’, the volatile profile of the mother plant was different from all its seedlings. Thus, the last emerging and weakest seedling is zygotic, while the rest are nucellar in polyembryonic mango genotypes. Volatile profiling is hence an effective biomarker to distinguish zygotic from nucellar seedlings in mango.

Toxic metals (TMs) are serious health hazard and are known to accumulate in fruits, which are consumed by people, owning to their distinct sweet flavor and potential health benefits. In the present study, arsenic (As), lead (Pb), chromium (Cr), copper (Cu), cadmium (Cd), nickel (Ni), and mercury (Hg) were measured by atomic absorption spectrometry (AAS) techniques in orange samples from 11 farmhouses of Sargodha, Pakistan. Principal component analysis (PCA) is a very useful analysis and data-reducing tool. This technique was used to explore the relationship between different metal levels in the oranges. The conventional wet acid digestion method was validated and assured with two (02) certified reference materials NIST-SRM 1515-Apple and (CRM) NIST-1570a Spinach Leaves by recovery test coupled with standard addition procedure. The results showed that Cd and Cr average daily intakes are 0.084 and 0.15 mg slightly higher than World Health Organization/Food and Agriculture Organization of the United Nations (WHO/FAO) recommended tolerable daily intake respectively. The assessment of health risk was also carried out in form of total hazard quotient (THQ) and hazard index (HI), indicating three heavy metals (Pb, Cd, and Cr) have significant human health risk potential to consumers while Cu, As, Hg, and Ni have non-significant (THQ < 1) health risks.