Desert Climate

The experiment was performed at the Grassland Ecological Research Station of Northeast Normal University, Jilin Province, China (44°40′-44°44′N, 123°44′-123°47′E). The research station has a semi-arid, continental climate with mean annual temperature ranged from 4.6 to 6.4°C (1950–2004). Mean annual precipitation ranged from 280 to 644 mm (1950–2014) with over 70% of the precipitation occurs from June to August. Potential evapotranspiration is approximately three times that of the annual precipitation. Vegetation is dominated by L. chinensis, a C₃ perennial rhizomatous grass; Phragmites australis, C. virgata and H. altissima are also abundant. Soil is classified as a chernozem soil, with 2.0% soil organic carbon content and 0.15% soil total nitrogen content (Wang et al., 2015 (link)).
One C₃ perennial grass (L. chinensis) and two C₄ grasses (annual: C. virgata; perennial: H. altissima) that co-occur in the meadow steppe of the study area were selected as experimental plants. L. chinensis is a widespread dominant grass of arid and semi-arid steppe in northern China, eastern Mongolia and Transbaikalia, Russia (Wang and Ba, 2008 (link)) and has an ability to resist drought, cold and alkaline conditions (Shi and Wang, 2005 (link)). C. virgata is widely distributed on the Northeast China Plain and is ecologically and economically important because of its high protein content and seed production. In addition, it also grows rapidly and is highly tolerant of alkaline conditions (Yang et al., 2008 (link); Lin et al., 2016 (link)). H. altissima is a perennial rhizomatous grass and is distributed in tropical, subtropical and temperate regions, especially in China and Southeast Asia. It has strong adaptability and stress resistance and can be used as a good soil and water conservation crop (Han et al., 2016 (link)).
On DOY 135 in 2013, seedlings of L. chinensis and H. altissima were transplanted to plastic pots (23.5 cm in diameter and 20 cm in height) filled with chernozem soil (8 kg soil pot⁻¹). For C. virgata, plants were germinated from seeds and transplanted to plastic pots. All species were planted as monocultures (five individuals per pot). Before the initiation of the drought treatment, all the transplanted plants were manually watered (to field capacity) every 3 days. To ensure that plant growth was not limited by nutrient elements, each pot received 2 mg of nitrogen fertilizer in the form of NH₄NO₃ every week. All the pots were watered thoroughly on the date (DOY 165) prior to the initiation of the drought treatment. During the drought experiment period (DOY 166–172), we stopped watering the plants. Moreover, all the pots were placed under a plastic shed to exclude natural precipitation. Variations in the soil water content for the studied grasses are provided in the supplementary information (Figure S1). The measurements of leaf gas exchange and collection of both leaf dark-respired CO₂ and fresh materials were conducted on DOY 166 (day 1 of the experiment), DOY 168 (day 3 of the experiment), DOY 170 (day 5 of the experiment) and DOY 172 (day 7 of the experiment). Before the initiation of the drought treatment, the studied grasses were in the stem elongation stage with average heights of 51.6, 62.9 and 42.4 cm for C. virgata, H. altissima and L. chinensis, respectively. The tiller densities of the studied grasses were 36 pot⁻¹, 19 pot⁻¹ and 17 pot⁻¹ for C. virgata, H. altissima and L. chinensis, respectively. Each pot had total leaf areas of 1,045 cm² pot⁻¹, 618 cm² pot⁻¹ and 648 cm² pot⁻¹ for C. virgata, H. altissima and L. chinensis, respectively.

Demography: This was a cross-sectional study among 1,006 patients with symptoms of allergic disorders who were referred to the allergy clinics of Mashhad University of Medical Science from October 2010 to February 2014. Patients without any apparent symptoms were excluded. All patients were from Khorasan, a vast area located in the Northeast of Iran with a cold semi-arid to hot-arid climate with a latitude of 36º N 59º E.
A demographic checklist including information such as age, gender, and patient history was completed, and on this basis, the exact date of the allergy was deduced in order to determine the seasonal distribution of symptomology (four groups). The two most commonplace methods for determine allergens, are the skin prick test (SPT), and the radio allegro sorbent test (RAST) in vitro and in vivo methods, respectively. As the SPT method is less time consuming and more cost effective, this was selected for our study.
SPT: The SPT was performed according to the patients’ history of aeroallergen hyper-sensitivity, with applying the standard allergen extract panel (GREER, USA) and compared with histamine chloride and normal saline, respectively, as positive and negative controls. Aeroallergen extracts for SPT included outdoor extracts including Russian thistle (Salsola kali) (G59), ash (Fraxinus excelsior) (P30), grass mix (TP27), tree mix (PO714), pigweed mix (P5) and lamb’s quarter (G43) and indoor extracts such as Dermatophagoides farina (D.farina) (B64), Dermatophagoides pteronyssinus (D. pteronyssinus) (B70), cockroach (BO12), Alternaria alternate (M1), Aspergillus mix (M04), penicillium (M05), cladosporium (M13), feather (E01),and cat fur(TE3). Choosing aeroallergens were based on Flora Iranica (4 ). In the SPT, a small drop of each allergen is placed on the volar surface of the forearm. A needle (25 gage) must touch each drop and penetrate into the epidermal surface at a low angle. The tip of the needle must then be gently lifted up to raise the epidermis without any bleeding, then allergen extracts should be dropped in the pricked area separately, then after about 15–20 minutes the solution is wiped away with a cotton tissue. The SPT shows a reaction which reaches a peak for allergens. The largest and smallest diameters of each reaction were measured; the average is usually reported for the result. A wheal diameter >3 mm and a flare diameter >10 mm are considered positive results (5 ).
Statistics: Statistical analyses were performed applying SPSS for Windows (version 22, New York, USA) as well as descriptive statistics and the Chi-square test. P<0.05 was considered statistically significant.

Study locations comprised 10 field stations situated in different climatic zones viz. coastal, arid, semi-arid, mountainous and subtropical (Fig. 1). Mosquitoes were collected from army cantonments from these field stations, with the permission of Director General Armed Forces Medical Services (DGAFMS), Ministry of Defence, Government of India. From each station, indoor resting adult Cx. quinquefasciatus females were collected from different sites at dawn and dusk. Collected mosquitoes were kept in pre-sterilized cages. Mosquitoes were anesthetized with chloroform and species was identified morphologically using standard taxonomic key. Adult mosquitoes were surface sterilized with 70% ethanol for 5 min followed by washing in phosphate buffered saline (PBS) (twice) before dissection for further processing of midgut isolates and bacterial cultivation according to Pidiyar et. al.[4] (link). Midguts were microscopically dissected out under sterile conditions and transferred individually to 100 µl of brain heart infusion broth (BHI broth) in 1.5 ml microcentrifuge tubes and enriched for 4 hrs at room temperature. After enrichment, an equal volume of 40% glycerol was added in each tube and stored in liquid nitrogen and transported to the laboratory for further study. From each station, 100 mosquitoes were dissected out and midgut samples were collected for bacterial diversity. To confirm the sterility of the procedure, two controls were also taken from each station which contained PBS from a mosquito's second wash.

This experiment was conducted at the experimental field of the Xinjiang Academy of Agricultural and Reclamation Sciences (longitude 86°03’E, latitude 44°19’N) from 2020 to 2021. The test area has an arid to semiarid continental climate. The average temperature during the growth period of rice was 17.7°C and 18.7°C, and the average rainfall was 0.3 mm and 0.4 mm in 2020 and 2021, respectively. Changes in precipitation and temperature during the rice growth period (May 1 to September 30) are shown in Figure 1. The experimental soil was loam in texture with an organic matter content of 11.21 g·kg^-1, total N of 0.74 g·kg^-1, alkaline N of 0.61 g·kg^-1, available P of 0.51 g·kg^-1, and available K of 93 g·kg^-1.