CaWRKY30 Positively Regulates Pepper Immunity by Targeting CaWRKY40 against Ralstonia solanacearum Inoculation through Modulating Defense-Related Genes.

The WRKY transcription factors (TFs) network is composed of WRKY TFs' subset, which performs a critical role in immunity regulation of plants. However, functions of WRKY TFs' network remain unclear, particularly in non-model plants such as pepper (Capsicum annuum L.). This study functionally characterized CaWRKY30-a member of group III Pepper WRKY protein-for immunity of pepper against Ralstonia solanacearum infection. The CaWRKY30 was detected in nucleus, and its transcriptional expression levels were significantly upregulated by R. solanacearum inoculation (RSI), and foliar application ethylene (ET), abscisic acid (ABA), and salicylic acid (SA). Virus induced gene silencing (VIGS) of CaWRKY30 amplified pepper's vulnerability to RSI. Additionally, the silencing of CaWRKY30 by VIGS compromised HR-like cell death triggered by RSI and downregulated defense-associated marker genes, like CaPR1, CaNPR1, CaDEF1, CaABR1, CaHIR1, and CaWRKY40. Conversely, transient over-expression of CaWRKY30 in pepper leaves instigated HR-like cell death and upregulated defense-related maker genes. Furthermore, transient over-expression of CaWRKY30 upregulated transcriptional levels of CaWRKY6, CaWRKY22, CaWRKY27, and CaWRKY40. On the other hand, transient over-expression of CaWRKY6, CaWRKY22, CaWRKY27, and CaWRKY40 upregulated transcriptional expression levels of CaWRKY30. The results recommend that newly characterized CaWRKY30 positively regulates pepper's immunity against Ralstonia attack, which is governed by synergistically mediated signaling by phytohormones like ET, ABA, and SA, and transcriptionally assimilating into WRKY TFs networks, consisting of CaWRKY6, CaWRKY22, CaWRKY27, and CaWRKY40. Collectively, our data will facilitate to explicate the underlying mechanism of crosstalk between pepper's immunity and response to RSI.

Molecular regulation of pepper innate immunity and stress tolerance: An overview of WRKY TFs.

The WRKY transcription factors (TFs) family constitutes a major group of TFs in spermatophytes. Different studies have endorsed the considerable biological roles performed by WRKY TFs in plant growth, biotic and abiotic stress responses. Genomic and transcriptomic profiling facilitate us in understanding the WRKY genes in various plants and reveal how WRKY TFs perform their action in response to different plant stresses. WRKY TFs actively take part in metabolism including carbohydrate synthesis, senescence, and secondary metabolites production. Molecular organization of WRKY TFs in plants highlight most predicted outcome of multiple responses simultaneously. Repression and activation related to W-box and other such elements is controlled at transcriptional, translational and domain level. WRKY TFs are becoming more important in crop improvement because of their binding with downstream elements. Additionally, WRKY proteins intermingle with various other TFs for modulating plant immunity. However, WRKY TFs self-regulation and crosstalk between different signaling pathways using WRKY TFs still need extensive investigations. In this review, we focused characteristics of WRKY TFs in Capsicum annum and related research advancement on their functional involvement in plant responses to the challenges of high temperature stress and pathogens infection. We summarized information about Capsicum annum WRKY TFs on the basis of their functions, their target genes and signaling pathways. Moreover, the mechanisms for synergistic responses to various biotic and abiotic stresses, WRKY target genes and other TFs as well will be of more interest with increments in existing information.

Influence of Industrial Wastewater Irrigation on Heavy Metal Content in Coriander (Coriandrum sativum L.): Ecological and Health Risk Assessment.

The primary objective of this study was to determine the heavy metal contents in the water-soil-coriander samples in an industrial wastewater irrigated area and to assess the health risks of these metals to consumers. Sampling was done from areas adjoining the Chistian sugar mill district Sargodha and two separate sites irrigated with groundwater (Site 1), and sugar mill effluents (Site 2) were checked for possible metal contamination. The water-soil-coriander continuum was tested for the presence of cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), manganese (Ni), lead (Pb), and zinc (Zn). The mean concentrations of all metals were higher than the permissible limits for all studied metals except for Mn in the sugar mill wastewater, with Fe (8.861 mg/L) and Zn (9.761 mg/L) exhibiting the highest values. The mean levels of Fe (4.023 mg/kg), Cd (2.101 mg/kg), Cr (2.135 mg/kg), Cu (2.180 mg/kg), and Ni (1.523 mg/kg) were high in the soil at Site 2 in comparison to the groundwater irrigated site where Fe (3.232 mg/kg) and Cd (1.845 mg/kg) manifested high elemental levels. For coriander specimens, only Cd had a higher mean level in both the groundwater (1.245 mg/kg) and the sugar mill wastewater (1.245 mg/kg) irrigated sites. An estimation of the pollution indices yielded a high risk from Cd (health risk index (HRI): 173.2), Zn (HRI: 7.012), Mn (HRI: 6.276), Fe (HRI: 1.709), Cu (HRI: 1.282), and Ni (HRI: 1.009), as all values are above 1.0 indicating a hazard to human health from consuming coriander irrigated with wastewater. Regular monitoring of vegetables irrigated with wastewater is strongly advised to reduce health hazards to people.

Bioaccumulation and transfer of zinc in soil plant and animal system: a health risk assessment for the grazing animals.

Heavy metals pollution has thorough worldwide apprehensions due to the instantaneous growth of industries. Farming regions are irrigated mainly with wastewater which contains both municipal and industrial emancipations. Keeping in view the above scenario, a study was designed in which three sites irrigated with ground, canal, and municipal wastewater in the District Jhang were selected to determine the zinc accumulation and its transfer in the soil, plant, and animal food chain. Zinc concentration was ranged as 18.85-35.59mg/kg in the soil, 26.42-42.67 mg/kg in the forage, and 0.982-2.85mg/kg in the animal samples. Investigated zinc concentration in soil and forages was found to be within the recommended WHO/FAO limits, but blood samples exceed the standards of NRC (2007). The maximum level of pollution load index (0.427-0.805mg/kg) and enrichment factor (0.373-0.894 mg/kg) for zinc was noticed upon wastewater irrigation. Daily intake (0.039 to 0.082 mg/kg/day) and health risk index (0.130 to 0.275 mg/kg/day) of zinc metal was higher in the buffaloes that feed on wastewater-irrigated forages. Bio-concentration factor (0.840 to 2.01mg/kg) for soil-forage was >1 which represents that these plants accumulated the zinc concentration into their tissues and raised health issues in grazing animals on consumption of wastewater-contaminated forages. As animal-derived products are part of human food, then zinc toxicity prevailed in livestock tissues ultimately affects the human food chain. Overall, findings of this study concluded that animal herds should be monitored periodically to devise preventive measures regarding the toxic level of heavy metals availability to livestock.

The impact of PEG-induced drought stress on seed germination and seedling growth of different bread wheat (Triticum aestivum L.) genotypes.

Wheat is an important crop, used as staple food in numerous countries around the world. However, wheat productivity is low in the developing world due to several biotic and abiotic stresses, particularly drought stress. Non-availability of drought-tolerant wheat genotypes at different growth stages is the major constraint in improving wheat productivity in the developing world. Therefore, screening/developing drought-tolerant genotypes at different growth stages could improve the productivity of wheat. This study assessed seed germination and seedling growth of eight wheat genotypes under polyethylene glycol (PEG)-induced stress. Two PEG-induced osmotic potentials (i.e., -0.6 and -1.2 MPa) were included in the study along with control (0 MPa). Wheat genotypes included in the study were 'KLR-16', 'B6', 'J10', '716', 'A12', 'Seher', 'KTDH-16', and 'J4'. Data relating to seed germination percentage, root and shoot length, fresh and dry weight of roots and shoot, root/shoot length ratio and chlorophyll content were recorded. The studied parameters were significantly altered by individual and interactive effects of genotypes and PEG-induced osmotic potentials. Seed germination and growth parameters were reduced by osmotic potentials; however, huge differences were noted among genotypes. A reduction of 32.83 to 53.50% was recorded in seed germination, 24.611 to 47.75% in root length, 37.83 to 53.72% in shoot length, and 53.35 to 65.16% in root fresh weight. The genotypes, 'J4', 'KLR-16' and 'KTDH-16', particularly 'J4' better tolerated increasing osmotic potentials compared to the rest of the genotypes included in the study. Principal component analysis segregated these genotypes from the rest of the genotypes included in the study indicated that these can be used in the future studies to improve the drought tolerance of wheat crop. The genotype 'J4' can be used as a breeding material to develop drought resistant wheat genotypes.

Effects of diverse irrigation with wastewater in soil and plants: assessing the risk of metal to the animal food chain.

In District Jhang, farmers use municipal wastewater to irrigate fodder crops as an alternative source to the deficient availability of fresh water. Therefore, the present study selected the three irrigation sources in District Jhang (canal water, ground water and municipal wastewater) to study the iron (Fe) concentration in the soil, fodder crops and ultimately their transfer into the animal body. Analysed Fe concentration varied as 16.40-27.53 mg/kg in soil samples, 19.72-30.34 mg/kg in fodder crops and 2.49-5.11 mg/kg in animals. Analysed Fe concentration in soil was higher on the wastewater irrigation site while canal water-irrigated fodder crop Zea mays exhibit the higher Fe concentration. In animal samples, higher Fe concentration was observed in the cow blood (4.09 mg/l), cow hairs (3.39 mg/kg) and cow faeces (5.11 mg/kg). Results of pollution load index (0.288-0.484 mg/kg) and enrichment factor (0.112-0.197 mg/kg) indicated that Fe concentration was minimally dispersed and enriched in these sites. Health risk and daily intake values were observed between the 0.029-0.059 and 0.042-0.084 mg/kg/day. Bio-concentration factor (0.834-1.47 mg/kg) for Fe which was greater than 1 explains that Fe contamination was transferred from the soil to fodder tissues and may raise health issues in the grazing animals if they are continuously exposed to these contaminated forages. Wastewater irrigation in study area has increased the Fe content in soil-plant environment that is a risking factor for animal and human health. Hence, this study recommended that wastewater should be treated prior to their irrigation on agricultural lands.

Field screening of diverse wheat germplasm for determining their adaptability to semi-arid climatic conditions.

Wheat (Triticum aestivum L.) is an important staple food crop for one third of global population and important crop for securing future food security. Rapid changes in the climate on global scale could be a threat for future food security. This situation urges plant breeders to explore the genetic potential of existing wheat germplasm. This study screened forty diverse wheat genotypes for their yield under two different agroclimatic conditions, i.e., Layyah and Dera Ghazi Khan, Pakistan. Data relating to plant height, peduncle length, flag leaf area, spike length, number of spikelets, number of grains per spike, thousand grain weight, chlorophyll content and grain yield were recorded. The tested wheat genotypes significantly differed for grain yield and related traits. Grain yield was positively correlated with plant height, spike length, spike number, flag leaf length, number of grains per spike, and 1000-grain weight. Biplot obtained from the cluster analysis by Euclidean method grouped the studied genotypes in 3 different groups. The genotypes exhibited 10.77% variability within quadrants, whereas 72.36% variability was recorded between quadrants according to clustering. Dendrogram grouped the tested genotypes into two main clusters. The main cluster "I" comprised of 2 genotypes, i.e., 'Seher-2006' and 'AS-2002'. The cluster "II" contained 38 genotypes based on Euclidian values. Genotypes within same cluster had smaller D2 values compared to those belonging to other clusters. The genetic relationships of genotypes provide useful information for breeding programs. Overall, the results revealed that genotypes 'Line 9733', 'Bhakar-2002', 'Line A9' and 'SYN-46' had better yield and yield stability under climatic conditions of southern Punjab. Therefore, these genotypes could be recommended for general cultivation in the study region.

Grain yield and correlated traits of bread wheat lines: Implications for yield improvement.

Global wheat yields are suffering due to differences in regional climatic conditions and soil fertility. Plant breeders are continuously working to improve the yield per unit area of wheat crop through selecting superior lines as parents. The screening and field evaluation of available lines allow the selection of superior ones and subsequently improved varieties. Therefore, heritable distinctions among 33 bread wheat lines for yield and related attributes were assessed under field conditions. The experiment included thirty lines and three check varieties. Data relating to different plant characteristics was collected at maturity. Significant differences were recorded for yield and related traits of tested wheat lines and check varieties. Wheat lines V6, V12 and V20 proved better with reduced number of days to reach anthesis and other desirable traits compared to check varieties. Days to start heading had strong correlation with spike length and number of spikelets spike-1. Flag leaf area had positive relationship with peduncle length and yield related traits. The 1000-garin weight and grain yield were also correlated with each other. It is concluded that V6, V10 and V20 proved better for all studied traits than the rest of the lines. Therefore, these lines could be used in wheat breeding program as parents to improve yield.

Deep placement of nitrogen fertilizer improves yield, nitrogen use efficiency and economic returns of transplanted fine rice.

Rice (Oryza sativa L.) feeds to two-third of the global population by serving as staple food. It is the main export commodity of several countries; thus, contributes towards foreign exchange earnings. Unfortunately, average global rice yield is far below than its genetic potential. Low nitrogen (N) use efficiency (NUE) is among the major reasons for low average yield. Current study evaluated the impact of nitrogen fertilizer application methods (conventional and deep placement) on growth, yield-related traits, chlorophyll contents, photosynthesis rate, agronomic N-use efficiency (ANUE), partial factors productivity of applied N (PFP) and economic returns of two different transplanted rice varieties (Basmati-515 and Super-Basmati). Fertilizer application methods significantly affected allometry, yield-related traits, chlorophyll contents, photosynthesis rate, ANUE, PFP and economic returns. Deep placement of N-fertilizer (DPNF) observed better allometric traits, high chlorophyll contents, photosynthesis rate, ANUE, PFP, yield attributes and economic returns compared to conventional application of N-fertilizer (CANF). Similarly, Basmati-515 had better allometric and yield-related traits, chlorophyll contents, photosynthesis rate, ANUE, PFP and economic returns than Super-Basmati. Regarding interactions among N-fertilizer application methods and rice varieties, Basmati-515 with DPNF resulted in higher chlorophyll contents, photosynthesis rate, ANUE, PFP, allometric and yield related traits and economic returns than CANF. The lowest values of these traits were observed for Super-Basmati with no application of N-fertilizer. Both varieties had better yield and economic returns with DPNF compared to CANF. It is concluded that DPNF improved yield, ANUE and economic returns; therefore, should be opted to improve productivity of transplanted fine rice. Nonetheless, lower nitrogen doses need to be tested for DPNF to infer whether it could lower N use in rice crop.

A study on the transfer of chromium from meadows to grazing livestock: an assessment of health risk.

The present investigation was performed in different district of Punjab to determine the chromium level in soil, forages, and blood plasma of the small ruminants (goat and sheep). The results showed that the mean concentrations of chromium in the soil of Sargodha, Mianwali, and Bhakkar districts were ranged from 1.8 to 2.3, 3.01 to 4.99, and 2.04 to 2.87 mg/kg, respectively. The chromium level was higher in Mianwali compared with Sargodha and Bhakkar. The mean concentrations of chromium in forages which were used for feeding purposes were found between 0.672 and 1.631, 1.493 and 2.612, and 0.7804 and 1.047 mg/kg for Sargodha, Mianwali, and Bhakkar, respectively. The mean concentrations of chromium in blood plasma of goat which consumed the contaminated forages were between 0.666 and 1.269 mg/L in Sargodha, 0.657 and 0.752 mg/L in Mianwali, and 1.39 and 2.37 mg/L in Bhakkar. In blood plasma of sheep, the residual levels of chromium in the districts Sargodha, Mianwali, and Bhakkar were ranged from 0.673 to 1.384, 0.83 to 1.086, and 1.496 to 3.611 mg/L, respectively. In the present research, there was a higher concentration of chromium in blood plasma of sheep from Bhakkar and the lowest was in Sargodha. The chromium level in blood samples was found to be higher than the tolerable level of 1.0 mg/L in all districts. In light of these results, it was concluded that chromium levels in the soil and forages of all sites were present within the safe limit while in blood samples of sheep and goats were found to be above the acceptable limit. Sheep and goats also consume water from river, streams, and ponds and other contaminated water sources. So it might be the reason of higher level of chromium in their blood plasma. Hence, the proper monitoring of study sites will be necessary to prevent the excessive accumulation of chromium in ruminants in the near future.

Correction to: A study on the transfer of chromium from meadows to grazing livestock: an assessment of health risk.

The affiliation of Shahzadi Mahpara is shown in this paper.

Insect pollinator diversity in four forested ecosystems of southern Punjab, Pakistan.

This study investigated pollinator assemblage diversity and richness in four forested ecosystems of southern Punjab, Pakistan, with different landscape types. Pirowal is situated in the plains of irrigated Punjab, Lal Suhanra is part of a sandy desert ecosystem, Ghazi Ghat is part of the Indus River delta, and Fort Munro is located in dry hilly mountains. A yearlong survey of pollinator populations was carried out in these four forested ecosystems from January to December of 2010. Fortnightly hand netting was performed for collecting flower-visiting insects whereas, pan traps of three colors (white, blue, and yellow) were deployed for collecting the data. A total of 8,812 individuals from two orders (Lepidoptera and Diptera) were observed, including 22 families and 154 species. Bees were the most abundant, with 4,502 individuals, and the most species-rich taxa, with 70 species in five families, followed by flies having 2,509 individuals and 51species in 10 families. Wasps were the least abundant with 1,801 individuals and 33 species in seven families. The assemblage structure of pollinator communities as visualized through rank abundance curves showed that there were many species with low abundance and only a few species with a much higher abundance. The most abundant species among the bees, in order, were Nomia sp.3, Megachile bicolor, and Colletes sp.3; among flies, Syrphus sp.2, Calliphoridae sp.1, and Empididae sp.4; and among wasps, Tiphiidae sp.1, Myzininae sp.2, and Scelionidae sp.1.