The Agrobacterium-mediated floral dip protocol is the most extensively used transformation method for a model plant Arabidopsis thaliana. Several useful methods for Agrobacterium tumefaciens–mediated transformations of Arabidopsis are existing, but they are time consuming and with low transformation efficiency. Here, we developed a transgenic Arabidopsis lines TET12p::TET12-RFP in a short period of time and enhanced transformation efficiency by using a modified transformation method by applying drought stress after floral dip. In this protocol, Agrobacterium cells carrying TET12p::TET12-RFP recombinant vectors were resuspended in a solution of 5% sucrose, 0.05% (v/v) silwet L-77 to transform female gametes of developing Arabidopsis inflorescences. Treated Arabidopsis were then applied with different levels of drought stresses to stimulate plants for the utilization of maximum plant energy in seed maturation process. The applied stresses achieved the fast maturation of already treated inflorescences while stopped the growing of newly arising untreated inflorescence, thus decreased the chances of wrong collection of untransformed seeds. Consequently, the collected seeds were mostly transgenic with a transformation frequency of at least 10%, thus the screening for positive transformants selection was more advantageous on a selective medium as compared to a classical floral dip method. Within 2-3 months, two hundred of individual transgenic plants were produced from just 10 infiltrated plants. This study concludes that application of drought stresses in a specific stage of plant is a beneficial strategy for achieving the transgenic Arabidopsis in a short period of time with high transformation efficiency.

Phenomic characterization through digital imaging (DI) can capture the three dimensional variation in wheat grain size and shape using different image orientations. Digital imaging may help identifying genomic regions controlling grain morphology using association mapping with simple sequence repeats (SSRs) markers. Accordingly, seed shape phenotypic data of a core collection of 55 wheat genotypes, previously characterized for osmotic and drought tolerance, were produced using computer based Smart grain software. Measured dimensions included seed volume, area, perimeters, length, width, length to width ratio, circularity, horizontal deviation from ellipse (HDEV), vertical deviation from ellipse (VDEV), factor form density (FFD) etc. The thousand grain weight (TGW) was positively correlated with grain size direct measurements; however, VDEV, FFD and other derived grain attributes showed no or negative correlation with TGW. Digital imaging divided the genotypes correctly into well-defined clusters. The wheat genotypes studied were further grouped into two sub-clusters by the Bayesian structure analysis using unlinked SSR markers. A number of loci over various chromosomal regions were found associated to grain morphology by the genome wide analysis using mixed linear model (MLM) approach. A considerable number of marker-trait associations (MTAs) on chromosomes 1D and 2D may carry new alleles with potential to enhance grain weight due to the use of untapped wild accessions of Aegilops tauschii. Conclusively, we demonstrated the application of multiple approaches including high throughput phenotyping using DI complemented with genome wide association studies to identify candidate genomic regions underlying these traits, which allows a better understanding on molecular genetics of wheat grain weight.

Human population growth and the accompanying increase in anthropogenic activities pose a significant threat to forest ecosystems by reducing the natural services these ecosystems provide. Malam Jabba, located in the District Swat of Pakistan’s Hindukush-Himalayan temperate zone, is known for its ecotourism, skiing, timber-producing tree species, medicinal plants, and unique biodiversity. However, a large portion of Swat Valley’s population depends on the Malam Jabba forests for timber and fuelwood. This study investigates how deforestation rates have increased in response to the growing human population in Malam Jabba, District Swat. To monitor forest cover changes, we used remote sensing (RS) and geographic information systems (GIS) tools. Vegetation analysis was conducted using the Normalized Difference Vegetation Index (NDVI) based on multi-temporal satellite imagery from 1980, 2000, and 2020. Using a decay model, we calculated the deforestation rate from 1980 to 2020 and projected future rates using MATLAB, based on anticipated population growth. Our results show that over the last two decades, the average annual deforestation rate rose from 0.7% to 1.93%, coinciding with a population increase from 1.2 million to 2.3 million at a growth rate of 9% per year. Projections indicate that the deforestation rate will increase to 2.5% annually over the next 20 years, given the predicted 11.6% yearly population growth. Population growth in District Swat has severely endangered nearby forest ecosystems, and further increases in human activity, such as unsustainable tourism, fuel and timber collection, and urbanization, will likely exacerbate this trend. Based on our findings, we recommend: (i) the implementation of reforestation programs and sustainable forest resource use; (ii) the development of a long-term forest management plan that maintains equilibrium between forest density and population pressure; and (iii) prioritizing areas with extreme human impact for in-situ conservation efforts.

Effective forest conservation and management needs to consider ecological factors and the susceptibility of different tree species to anthropogenic activity. Dry deciduous forests in the tropics have been undervalued because of their low timber production compared to temperate and boreal forests. In order to quantify the current significance of Pakistan’s tropical dry deciduous forests in relation to ecological variables and anthropogenic threats, a broad phytosociological survey was conducted. In the study area, a total of 140 species of woody plants belonging to 52 families were identified. The cluster analysis depicts the distribution of nine plant communities in five clusters, in which Dodonaea viscosa-Acacia modesta-Dulbergia sissoo community was the most dominant community, showing cosmopolitan distribution in the study area. The results are further authenticated by DCA and CCA analyses, which indicate that altitude, precipitation, and temperature are the most important factors influencing the distribution and composition of tropical dry deciduous forests along the Himalayan foothills. In the study area, an annual fuelwood consumption of 270.38 tons was recorded. It was discovered that high-altitude forests were subject to extensive tree harvesting, overgrazing, browsing, and high fuelwood consumption. In addition, these forests have a low regeneration rate from 4.5 to 4.8 seedlings/quadrat, due to extensive human activities. Remarkable differences in the anthropogenic pressure and disturbances were found in the protected and unprotected forests. Based on our findings, we recommend that: (i) Immediate management intervention and an in-situ conservation strategy must be implemented in areas exposed to high levels of anthropogenic threats. (ii) Unsustainable grazing and fuelwood collection must be managed, and high-threat areas must be immediately prohibited. (iii) The local populace must be made aware of the grave consequences of anthropogenic disturbances, and a collaborative management strategy must be implemented.

Human population growth and associated increases in anthropogenic activities pose a significant threat to forest ecosystems by diminishing the natural ecosystem services these systems provide. Malam Jabba is located in District Swat Pakistan’s Hindukush-Himalayan temperate zone, which is renowned for ecotourism and skiing and is rich in timber-producing tree species, medicinal plants, and unique biodiversity; however, the majority of Swat Valley’s population relies on Malam Jabba forests for their timber & fuelwood requirements. We examined how the deforestation rate increased with increasing human population density in Pakistan’s Malam Jabba area of the Hindukush-Himalayas. To identify the forest cover, remote sensing, and geographic information systems were used (RS & GIS). The study area’s vegetation was analyzed with the Normalized Divergence Vegetation Index (NDVI) using multitemporal satellite images for the years 1980, 2000, and 2020. The deforestation rate from 1980 to 2020 was then determined using the decay model, and the MATLAB program was used to predict the deforestation rate for the following two decades in relation to the anticipated growth in the human population. Our result revealed that, in the last two decades, the average rate of deforestation increased from 0.7% to 1.93% per year, while the human population of District Swat increased from 1.2 to 2.3 million at a rate of 9% per year. The decay model predicts that the study area’s deforestation rate will increase to 2.5% per year over the next two decades due to the forecasted 11.6% per year population growth rate. Human population growth in District Swat, Pakistan has seriously threatened the nearby forest ecosystems, and a future increase in human population will further accelerate anthropogenic activities like unsustanible tourism, fuel and timber wood collection and urbanization. Based on our results, we recommend that: (i) in addition to reforestation programs and sustainable use of forest resources, the government should implement a long-term forest management plan (ii) where the density of forest cover can be sustained at an equilibrium level dependent of population growth pressure (iii) and areas with extreme human pressure should be designated as most important for in situ conservation approach.

The trade of pinus gerardiana nuts (Chilghoza) is critical for the economy and livelihoods of communities in District North Waziristan Agency, formerly FATA. Its commercial value supports local economies and provides employment, highlighting the importance of sustainable management and market access for these valuable non-timber forest products. This research investigates the effects of militancy on non-timber forest products (NTFPs) trade, particularly Chalghoza nuts, in District North Waziristan Agency, previously known as Federally Administered Tribal Areas (FATA). NTFPs play a crucial role in the livelihoods and economies of many communities globally. However, regions affected by militancy often face challenges disrupting NTFP trade, impacting local populations' livelihoods. The study employs quantitative analysis and qualitative case studies to explore the relationship between militancy and NTFPs, focusing on Chalghoza nuts. Data were collected from key stakeholders, including NTFPs and Chalghoza nuts collectors, traders, and government officials, during and after periods affected by terrorism and military operations. Twenty-seven plant species were identified as commercially important NTFPs, with Chalghoza nuts being the most significant due to their commercial value and support to the local economy. Findings indicate that women and youth predominantly collect NTFPs and Chalghoza nuts, while the market trade is controlled by older men. Most respondents were illiterate and untrained, relying on personal experiences and traditional methods for collection, transportation, and packing. The study highlights significant impacts of terrorism and military operations on Chalghoza nuts trade in North Waziristan Agency, FATA. The presence of terrorism creates instability, fear, and insecurity, leading to 32% unemployment among Chalghoza collectors/traders, with 14% shifting to other sectors. An inequitable profit gap was recorded among local collectors, middlemen, and traders, sometimes exceeding 200%. Military operations and militancy disrupt production, transportation, and market access, resulting in decreased productivity, increased costs, and reduced profit. Additionally, military operations cause collateral damage, casualties, and destruction of Chalghoza market infrastructure and supply chains, exacerbating industry challenges. The study recommends conflict-sensitive resource management strategies prioritizing NTFP-dependent communities' security and welfare, ensuring safe access, sustainable harvesting, and equitable trade to uplift the rural economy.