DNA barcodes were obtained for 81 butterfly species belonging to 52 genera from sites in north-central Pakistan to test the utility of barcoding for their identification and to gain a better understanding of regional barcode variation. These species represent 25% of the butterfly fauna of Pakistan and belong to five families, although the Nymphalidae were dominant, comprising 38% of the total specimens. Barcode analysis showed that maximum conspecific divergence was 1.6%, while there was 1.7–14.3% divergence from the nearest neighbour species. Barcode records for 55 species showed <2% sequence divergence to records in the Barcode of Life Data Systems (BOLD), but only 26 of these cases involved specimens from neighbouring India and Central Asia. Analysis revealed that most species showed little incremental sequence variation when specimens from other regions were considered, but a threefold increase was noted in a few cases. There was a clear gap between maximum intraspecific and minimum nearest neighbour distance for all 81 species. Neighbour-joining cluster analysis showed that members of each species formed a monophyletic cluster with strong bootstrap support. The barcode results revealed two provisional species that could not be clearly linked to known taxa, while 24 other species gained their first coverage. Future work should extend the barcode reference library to include all butterfly species from Pakistan as well as neighbouring countries to gain a better understanding of regional variation in barcode sequences in this topographically and climatically complex region.

Sequences from the DNA barcode region of the mitochondrial COI gene are an effective tool for specimen identification and for the discovery of new species. The Barcode of Life Data Systems (BOLD) (www.boldsystems.org) currently hosts 4.5 million records from animals which have been assigned to more than 490,000 different Barcode Index Numbers (BINs), which serve as a proxy for species. Because a fourth of these BINs derive from Lepidoptera, BOLD has a strong capability to both identify specimens in this order and to support studies of faunal overlap. DNA barcode sequences were obtained from 4503 moths from 329 sites across Pakistan, specimens that represented 981 BINs from 52 families. Among 379 species with a Linnaean name assignment, all were represented by a single BIN excepting five species that showed a BIN split. Less than half (44%) of the 981 BINs had counterparts in other countries; the remaining BINs were unique to Pakistan. Another 218 BINs of Lepidoptera from Pakistan were coupled with the 981 from this study before being compared with all 116,768 BINs for this order. As expected, faunal overlap was highest with India (21%), Sri Lanka (21%), United Arab Emirates (20%) and with other Asian nations (2.1%), but it was very low with other continents including Africa (0.6%), Europe (1.3%), Australia (0.6%), Oceania (1.0%), North America (0.1%), and South America (0.1%). This study indicates the way in which DNA barcoding facilitates measures of faunal overlap even when taxa have not been assigned to a Linnean species.

Plant growth regulators and Rhizobium are actively involved in the regulation of flowering, pod formation, nodulation, and ultimately the growth and yield of legumes. However, very limited information is available on the combined effect of gibberellic acid (GA3) and Rhizobium on growth attributes and yield of legume crops. This experiment was designed to fill this gap by studying the performance of chickpea under exogenous application of GA3 (10−4 and 10−5 M) alone and in combination with Rhizobium. Exogenous application of GA3 (10−5 M) combined with rhizobium inoculation gave the highest values for number of nodules per plant (16) and their dry biomass (0.22 g). Moreover, GA3 application and seed inoculation with Rhizobium, when applied singly, significantly enhanced chickpea growth. However, the most promising results were obtained by the inoculation of Rhizobium accompanied with GA3 (10−5 M). Plant height, grain and stover yield, and chlorophyll contents were enhanced up to 35%, 39%, 21%, and 51%, respectively. Likewise, the bioaccumulation of macronutrients (N, P and K) was maximum in plants receiving both Rhizobium inoculation and GA3 application. It is concluded that the combined application of Rhizobium and GA3 has synergistic effects on the growth, yield, and nutrient contents of chickpea.

Morphological study of 1,795 spiders from sites across Pakistan placed these specimens in 27 families and 202 putative species. COI sequences >400 bp recovered from 1,782 specimens were analyzed using neighbor-joining trees, Bayesian inference, barcode gap, and Barcode Index Numbers (BINs). Specimens of 109 morphological species were assigned to 123 BINs with ten species showing BIN splits, while 93 interim species included representatives of 98 BINs. Maximum conspecific divergences ranged from 0–5.3% while congeneric distances varied from 2.8–23.2%. Excepting one species pair ( Oxyopes azhari – Oxyopes oryzae ), the maximum intraspecific distance was always less than the nearest-neighbor (NN) distance. Intraspecific divergence values were not significantly correlated with geographic distance. Most (75%) BINs detected in this study were new to science, while those shared with other nations mainly derived from India. The discovery of many new, potentially endemic species and the low level of BIN overlap with other nations highlight the importance of constructing regional DNA barcode reference libraries.

Cotton (Gossypium hirsutum ssp.) belongs to the genus Gossypium from the family Malvaceae. About 100 genera are in the family Malvaceae and about 1500 species. It is grown worldwide for oil, seed, and fiber. A major problem in cotton cultivation is a drought that’s why this experiment was planned to evaluate the cotton mutants for drought tolerance. A variety of cotton Cyto-155 was mutated with gamma rays at different intensities such that 20kR, 25kR, 30kR, and 35kR, to obtain some genetic variation. Mutated seed along with non-mutated of that cotton variety was sown in split plot layout under randomized complete block design with three replications, under the normal irrigated condition with six irrigations and water stress condition with two irrigations at maturity data was collected for plant height, number of monopodial branches, number of sympodial branches, bolls per plant, boll weight, seed index, GOT%, lint index, seed cotton yield, relative water content and excised leaf water loss.Results showed a highly significant effect of genotype and water regime on all observed traits. Water regime × genotype interaction was also highly significant for plant height, number of sympodial branches, seed index, boll weight, lint index, and excised leaf water loss. Water regime × genotype interaction was non-significant for number of monopodial branches, bolls per plant, GOT%, seed cotton yield, and relative water content. Mean comparison indicated that seed mutated at 30kR and 35kR performed well for all yield-related traits in both irrigated conditions.

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For growth or development of pulses, biotic and abiotic environmental factors are more conspicuous under stress conditions. For the survival against abiotic stresses, salicylic acid (SA) is reported a universal remedy. At the Soil Bacteriology Section, Ayub Agricultural Research Institute, Faisalabad, a pot study was conducted to monitor the role of Plant Growth Promoting Rhizobacteria (PGPR) and Salicylic acid in chickpea under salt stress. Eight treatments including control PGPR inoculation and Salicylic acid with their different combination were used. Results revealed that positive response of PGPR on productivity of chickpea but more enunciated response about grain yield was observed with the combined application of SA and PGPR compared to control. Growth parameters i.e root length, root mass, number of nodules and shoot mass were highly affected where SA was applied along with PGPR. From the study, it is proposed that under salt stress the combination of SA + PGPR can be a suitable practice for more production of chickpean Pakistan.

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