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    Environment is associated with chytrid infection and skin microbiome richness on an amphibian rich island (Taiwan)

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    Evolutionary implications of new Postopsyllidiidae from mid-Cretaceous amber from Myanmar and sternorrhynchan nymphal conservatism

    Systematic palaeontologyOrder Hemiptera Linnaeus, 1758Suborder Sternorrhyncha Amyot et Audinet-Serville, 1843Superfamily Protopsyllidioidea Carpenter, 1931Family Postopsyllidiidae Hakim, Azar et Huang, 2019Genus Megalophthallidion Drohojowska et Szwedo, gen. nov.LSID urn:lsid:zoobank.org:act:A6F71390-9B8E-4A19-8F30-C2A024B6EFB1Type speciesMegalophthallidion burmapateron Drohojowska et Szwedo, sp. nov.; by present designation and monotypy.EtymologyGeneric name is derived from Classic Greek megas (μέγας)—large, ophthalmos (ὀφθαλμός)—an eye and Greek form of generic name Psyllidium. Gender: masculine.Type localityNorthern Myanmar: state of Kachin, Noije bum 2001 Summit Site amber mine in the Hukawng Valley, SW of Maingkhwan.Type stratumLowermost Cenomanian, Upper Cretaceous (‘mid-Cretaceous’).DiagnosisHead capsule with 12 stiff setae on tubercles (18 setae in Postopsyllidium); fore wing without pterostigma (tiny pterostigma, widening of ScP + RA present in Postopsyllidium); vein CuP not thickened distally (distinctly thickened distally in Postopsyllidium); profemur with a row of ventral (ventrolateral) setae (two rows in Postopsyllidium).Megalophthallidion burmapateron Drohojowska et Szwedo, sp. nov.LSID urn:lsid:zoobank.org:act:F3F971F4-AE04-4F41-98B0-9A0A04470625.(Figs. 1A–F, 2A–I).Figure 1Megalophthallidion burmapteron gen. et sp. nov., holotype (MAIG 6687), imago. (A) Photo of body, ventral side; (B) photo of right antennae and (C) drawing of antenna; (D) drawing of body, dorsal side; (E) drawing of thorax structure with sclerites marked: red—pronotum; orange—mesopraescutum; yellow—mesoscutum; light green—mesoscutellum, dark green—mesopostnotum; light blue—metascutum; dark blue—metascutellum; violet—metapostnotum; (F) photo of thorax dorsal side. Scale bars: 0.5 mm (A), 0.2 mm (B–D), 0.1 mm (F).Full size imageFigure 2Megalophthallidion burmapteron gen. et sp. nov., holotype (MAIG 6687), imago. (A) Photo of right fore wing; (B) photo of right wings; (C) photo of antenna and proleg; (D) photo of proleg and mesoleg, and (E) photo of femur of proleg, and (F) photo of right metatarsus and left mesotarsus in the background, and (G) photo of right mesotarsus of mesoleg, and (H) Photo of tarsi; (I) photo of male genital block. Scale bars: 0.5 mm (A–D), 0.2 mm (B,E,F,H), 0.1 mm (G,I).Full size imageMaterialHolotype, number MAIG 6687 (BUB 96), deposited in Museum of Amber Inclusions (MAIG), University of Gdańsk, Poland. Imago, a complete and well-preserved male. Piece of amber 8 × 6 × 3 mm, cut from larger lump, polished flat on both sides.Type localityNorthern Myanmar: state of Kachin, Noije bum 2001 Summit Site amber mine in the Hukawng Valley, SW of Maingkhwan.Type stratumLowermost Cenomanian, Upper Cretaceous (‘mid-Cretaceous’).DiagnosisAs for the genus with the following additions: three ocelli distinct, antennomere IX the longest, about as long as pedicel, antennomeres III–VII and XI of similar length, antennomere XII the shortest, subconically tapered in apical portion. Paramere lobate, ventral margin with acute, small process, apical and dorsal margins rounded. Aedeagus geniculately bent at base, directed dorsally, tapered apicad.DescriptionMale (Figs. 1A–F, 2A–I). Head with compound eyes distinctly wider than pronotum (Fig. 1D–F). Compound eyes subglobular, protruding laterally. Vertex short in midline, about 2.5 times as wide as posterior margin and as long in middle; trapezoidal, anterior margin slightly arched, lateral margins diverging posteriad, posterior margin shallowly arched, disc of vertex with distinct setae on large tubercles: four setae at posterior margin, two at anterior angles of compound eyes, two medial, over the median ocellus. Three ocelli present, median ocellus distinct, visible from above, lateral ocelli near anterior angles of compound eyes. Frons about as wide as long in midline, two rows of setae on tubercles, upper row at level of median ocellus, lower one, below half of compound eye height. Clypeus, elongate, triangular, in lower portion roof-like; two setae on tubercles near upper margin. Genae very narrow. Rostrum reaching slightly beyond mesocoxae, apical segment slightly shorter than subapical one, darker. Antennae bases placed at lower margin of compound eyes; antennal fovea elevated; scapus shorter than pedicel, cylindrical; pedicel cylindrical; antennomeres IIIrd–VIIth and XIth of similar length, VIIIth slightly longer than VIIth, as long as Xth antennomere, IXth the longest, XIIth the shortest, tapered apically; rhinaria absent.Thorax (Fig. 1D–F): pronotum quadrangular, about as long as mesothorax; pronotum with anterior and posterior margins parallel, merely arcuate, disc with transverse groove in the median portion, lateral margins slightly arcuate, two distinct setae on tubercles in anterolateral angle, two setae on tubercles anterior margin at distance1/3 to median line, three distinct setae on tubercles in posterolateral angles. Mesopraescutum subtriangular, with apex widely rounded, about 0.4 times as wide as pronotum, about 0.4 times as long as wide, delicately separated from mesoscutum. Mesoscutum as wide as pronotum at widest point, distinctly narrowed medially, anterior angles rounded, anterolateral margin sigmoid, lateral angle acute, posterior angles wide, posterior margin V-shape incised, posterolateral areas of mesoscutum disc declivent posteriorly; disc with two setae on tubercles, at 1/3 of mesoscutum width. Mesoscutellum about as long as wide, diamond-shape, anterior and lateral angles acute, posterior angle rounded. Mesopostnotum in form of transverse band, slightly widened in median portion. Metascutum narrower than mesoscutum, anterior angles widely rounded, lateral angles acute, anterolateral margin concave, posterior margin arcuate, with deep median arcuate incision. The suture between metascutum and metascutellum weakly visible, metascutellum subtriangular, longer than wide at base.Parapteron with three distinct setae.Fore wing (Fig. 2A,B) membranous, narrow, elongate, about 3.5 times as long as wide, widest at 2/3 of length. Anterior margin merely arcuate, slightly bent at very base, anteroapical angle widely arcuate, apex rounded, posteroapical angle widely arcuate, tornus arcuate, claval margin straight, with incision between terminals of Pcu (claval apex) and A1. Stem ScP + R + MP + CuA slightly arcuate, very short stalk ScP + R + MP + CuA leaving basal cell, stem ScP + R oblique, straight, forked in basal half of fore wing length, branch ScP + RA, oblique, reaching anterior margin slightly distally of half of fore wing length, slightly distally of ending of CuA2 branch; branch RP slightly arcuate, a little more curved in basal section, reaching margin at anteroapical angle; stalk MP + CuA slightly shorter than basal cell; stem MP almost straight, forked in apical half of fore wing, at about 2/3 of fore wing length, with three terminals reaching margin between apex and posteroapical angle; stem CuA shorter than branches CuA1 and CuA2, about half as long as branch CuA1; claval vein CuP weak at base, not thickened distally; claval vein Pcu straight, claval vein A1 straight. Basal cell present, subtriangular, about twice as long as wide, basal veinlet cua-cup oblique, no other veinlets present; cell r (radial) very long, longer than half of fore wing length; cell m (medial) the shortest, shorter than cell cu (areola postica). Margins of fore wing with fringe of long setae, starting on costal margin near base of fore wing, ending at level of middle of cell cu; longitudinal veins with distinct, scarcely but evenly dispersed, movable setae; terminal section of CuP with two setae; costal margin with row of short, densely distributed setae, apical margin, tornus and claval margin with rows of scaly setae.Hind wing (Fig. 2B) membranous, shorter than fore wing, 3.23 times as long as wide. Costal margin bent at base, then almost straight up to the level of ScP + RA end and wing coupling lobe, then straight to anteroapical angle, anteroapical angle widely arcuate, apex arcuate, posteroapical angle arcuate, tornus straight, claval margin merely arcuate, posteroclaval angle angulate; stem ScP + R + MP bent at base, then straight, stem ScP + R short, branch ScP + RA short, about as long as stem ScP + R, branch RP arcuate basally than straight, reaching apex; stem MP arcuate, forked slightly distad CuA1 terminus level, branch MP1+2 slightly arcuate, reaching margin at posteroapical angle, branch MP3+4 straight, reaching tornus; stem CuA slightly bent at base, then straight, forked slightly distad ScP + R forking, branch CuA1 arcuate, branch CuA2 short, straight, slightly oblique, reaching tornus; claval vein CuP weak, visible only at base, claval vein Pcu slightly arcuate; wing coupling apparatus (fold) with a few short setae.Legs slender, relatively long, profemora armed (Fig. 2C–H). Procoxa as long as profemur, narrow, flattened. Protrochanter scaphoid, elongate, with long apical and subapical setae. Profemur flattened laterally, about as long as protibia, ventrally armed with four large setae on elevated plinths; dorsal margin with row of short, decumbent setae. Protibia narrow, rounded in cross section, covered with short setae, a few longer setae in distal portion. Protarsus—single, long tarsomere, plantar surface with row of semi-erect setae; tarsal claws long, straight, directed ventrally, no arolium nor empodium.Mesocoxa elongate, narrow, slightly flattened. Mesotrochanter scaphoid. Mesofemur slender, flattened laterally, dorsal margin with short setae. Mesotibia subequal to mesofemur, slender, covered with setae, two apical setae slightly thicker and longer. Mesotarsus with three tarsomeres, basimesotarsomere the longest, shorter than cumulative length of mid- and apical mesotarsomere, plantar margins with setae, two apical setae slightly longer and thicker; midmesotarsomere the shortest, 1/3 of basimesotarsomere length, a few setae on plantar surface; apical tarsomere shorter than basimesotarsomere, twice as long as midmesotarsomere, plantar surface with a few, scarcely dispersed setae, tarsal claws long, narrow, directed ventrally, no arolium nor empodium.Metacoxa conical, narrow. Metatrochanter scaphoid, elongate. Metafemur slender, laterally flattened, longer than mesofemur, dorsal margin with row of short setae. Metatibia, long, slender, 1.6 times as long as metafemur, with suberect setae of different size, two larger and longer and two shorter setae subapical setae. Metatarsus slightly less than half of metatibia length, with three tarsomeres, basimetatarsomere the longest, more than twice as long as apical metatarsomere, 1.5 times as long as combined length of mid- and apical metatarsomere, plantar surface with scarce decumbent setae; mid metatarsomere the shortest, 1/4 of basimetatarsomere length, plantar surface with a few setae, two apical ones slightly thicker; apical metatarsomere about 0.4 of basimetatarsomere length, with scarcely dispersed setae on along plantar surface; tarsal claws, long, slender, other pretarsal structures absent.Abdomen (Fig. 1F) narrowly attached to thorax, tergite segment shorter, 2nd tergite distinctly longer, 3rd to 8th tergites of similar length; pygofer narrowing apicad, ventral margin strongly elongated posteriorly; anal tube short, directed posterodorsad, anal style shorter than anal tube. Paramere lobate, ventral margin with acute, small process, apical and dorsal margins rounded. Aedeagus (Fig. 2I) geniculately bent at base, directed dorsad, tapered apicad.Female. Unknown.Megalophthallidion sp. (5th instar nymph)(Figs. 3A–D, 4A–F)Figure 3Megalophthallidion sp. (MAIG 6688), nymph. (A) Photo of body, dorsal side and (B) drawing of body dorsal side; (C) photo of body dorsal side and (D) drawing of body ventral side. Scale bars: 0.5 mm (A–D).Full size imageFigure 4Megalophthallidion sp. (MAIG 6688), nymph. Photo of clypeus and (B) drawing of clypeus; (C) photo of proleg, and (D) photo of mesoleg, and (E) photo of metaleg; (F) photo of posterior part of abdomen ventral side. Scale bars: 0.1 mm (A–F).Full size imageMaterialNymph, 5th instar, MAIG 6688 (BUB 1799), deposited in Museum of Amber Inclusions (MAIG), University of Gdańsk, Poland. Piece of amber 13 × 6 × 2 mm, cut from larger lump, polished flat on one side, more convex on the other.Diagnostic charactersThe nymph of Megalophthallidion gen. nov. is similar in general body shape to the only known fossil protopsyllidioidean nymph described from Lower Cretaceous Lebanese amber—Talaya batraba Drohojowska et Szwedo, 2013. The nymph of Talaya batraba is 2nd or 3rd instar, therefore some features are difficult to compare with this last instar nymph of Megalophthallidion gen. nov. The morphological states observed in those two specimens are: head covered with strongly expanded disc and expanded disc of pronotum, however shapes and ratios of these structures differ; compound eyes on ventral side of head, shifted laterad (ommatidia on cones in T. batraba, while ventroposterior expansions are present in Megalophthallidion gen. nov.); compound eyes visible from above as short, stout cones in fissure between posterior margin of disc (hypertrophied vertex) and anterior margin of pronotum (compound eyes (?) are visible on dorsal side of Permian Aleuronympha bibulla Riek, 1974); in Megalophthallidion gen. nov. rostrum reached mesocoxa, while in Talaya batraba distinctly exceeds length of the body; abdomen with 9 segments; tergites of abdominal segments 5th–9th expanded posterolaterad in form of fan-like expansion; 9th abdominal segment short, placed ventral; anal tube short, cylindrical, epiproct (?) globular.DescriptionNymph, 5th instar (Figs. 3A–D, 4A–F). Body oval shaped, dorso-ventrally flattened, 1.5 times longer than wide with segmentation visible; on the ventral side slightly concave. Length of body c. 1.56 mm long, outline, in dorsal view, maximum width of body 0.94 mm; length of head and pronotum (cephaloprothorax) c. 0.46 mm in midline, width c. 0.83 mm; cumulative length of mesonotum + metanotum c. 0.25 mm; abdomen c. 0.8 mm long. Dorsal side (Fig. 3A,B) with distinct median line (ecdysial line), not reaching anterior or posterior margin of the body, the line distinctly roof-like in abdominal portion. Anterior margin of head (cephaloprothorax) disc arcuate, lateral angles rounded; anterior margin of pronotum arcuate, lateral margins arcuately diverging posteriad, posterior margin distinctly arcuate, anterior angles widely rounded, posterior angles acutely rounded, disc elevated, convex, lateral portions declivitous; the fissure between posterior margin of head disc and anterior margin of pronotum narrow, widened medially, with stalked compound eyes popping out.Head partly separated from prothorax, wide in ventral view. Bases of antennae protruding anterolaterally, wide, anterior margin arcuate, with a small lump extending anteriorly connecting margin with vertex expansion. Suture separating anteclypeus and postclypeus visible in ventral aspect (Fig. 4A,B). Postclypeus about three times as long as wide, oval, slightly swollen, without any setae; weak traces of salivary pump muscle attachments visible. Anteclypeus about as long as postclypeus, widened in upper section below clypeal suture, convex, carinately elevated in lower section, with sides distinctly declivitous, clypellus long, carinately elevated. Lora (mandibulary plates) distinct, separated from anteclypeus by shallow suture, with upper angles at half of postclypeus length, lower angles at half of anteclypeus length, about as wide as half of postclypeus width. Maxillary plates narrow. Genal portion of head enlarged, medial portion arcuately convex; lateral sections narrowing laterally, terminally encircling bases of compound eyes. Antennae short (Fig. 3C,D), placed in front of genal portion. Antennal flagellum indistinctly subdivided into four segments. Rostrum (Fig. 4A,B) three-segmented, 0.2 mm long, with apex reaching apex of mesocoxae; apical segment about 2.5 times as long as subapical one.No lateral sclerites on meso- and metathorax, only one plus one large medial sclerite on both meso- and metathorax. Mesothoracic and metathoracic wing pads distinct, wide, subtriangular, with posterior apices directed posteriorly; lateral portions of mesothoracic wing pads arcuate. Fore wing pad 0.6 mm long, with small, straight humeral lobe, forming a right angle, not protruding anteriorly. Mesothoracic tergites slightly larger than metathoracic segments (respectively c. 0.14 mm and c. 0.12 mm long in midline, 0.26 mm and 0.27 mm in lateral lines); mesothoracic tergum with distinct median elevation (low double crest with ecdysial line in between), slightly wider than long in midline, anterior margin arcuate, lateral margins straight, subparallel, posterior margin concave. Metathoracic wing pad apex slightly exceeding mesothoracic wing pad. Metathoracic tergum wider than long, slightly shorter than mesothoracic tergum, with distinct elevation in the middle.Legs relatively long (Figs. 3C,D, 4C–E). Coxae of legs placed near the median axis of the body. Prolegs: procoxal pit with margins elevated, procoxa conical (c. 0.1 mm long), protrochanter scaphoid, about as long as procoxa, profemur c. 0.13 mm long, slightly flattened laterally, merely thickened, protibia longer than profemur, c. 0.23 mm long; tarsus shorter than protibia, basiprotarsomere about as long as apical protarsomere, the latter with distinct tarsal claws, and wide arolium. Mesoleg similar to proleg, mesocoxa conical (c. 0.1 mm long), mesotrochanter scaphoid, mesofemur (c. 0.13 mm) slightly flattened laterally, mesotibia slightly longer than mesofemur (c. 0.18 mm), mesotarsus slightly shorter than mesotibia, three-segmented, basimesotarsomere the longest (c. 0.07 mm), about as long as combined length of mid- and apical mesotarsomeres (c. 0.04 mm respectively), arolium wide, tarsal claws distinct. Metaleg: metacoxa conical (c. 0.1 mm), metatrochanter scaphoid, about as long as metacoxa (c. 0.12 mm). Metafemur (c. 0.17 mm) slightly more thickened than pro- and mesofemur, metatibia slightly longer (0.19 mm) than pro- and mesotibiae. Metatarsus three-segmented: basimetatarsomere about as long (0.08 mm) as combined length of mid- and apical metatarsomeres (0.04 mm respectively), arolium lobate, wide, tarsal claws distinct, widely spread.Abdomen (Fig. 3A–D) 9-segmented, narrow at base, widening fan-shape posteriorly, 1st segment visible from above, segmentation visible, abdominal terga 5th–9th expanded posterolaterally. Tergites carinately elevated in the middle, separated by ecdysial line. 1st sternite visible in ventral view, sternites 2nd–4th fused medially, sternites 5th–9th separated; 9th abdominal segment short (Fig. 4F), placed ventrally, under tergal expansion; anal tube short, cylindrical, epiproct (?) globular. More

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    Carcass detection and consumption by facultative scavengers in forest ecosystem highlights the value of their ecosystem services

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    General description of the experimental areaThe experiment was performed for two years at the National Key Irrigation Experimental Station located on the Songnen Plain in Heping town, Qing’an County, Suihua, Heilongjiang, China, with a geographical location of 45° 63′ N and 125° 44′ E at an elevation of 450 m above sea level (Fig. 1). This region consists of plain topography and has a semiarid cold temperate continental monsoon climate, i.e., a typical cold region with a black soil distribution area. The average annual temperature is 2.5 °C, the average annual precipitation is 550 mm, the precipitation is concentrated from June to September of each year, and the average annual surface evaporation is 750 mm. The growth period of crops is 156–171 days, and there is a frost-free period of approximately 128 days year−122. The soil at the study site is albic paddy soil with a mean bulk density of 1.01 g/cm3 and a porosity of 61.8% prevails. The basic physicochemical properties of the soil were as follows: the mass ratio of organic matter was 41.8 g/kg, pH value was 6.45, total nitrogen mass ratio was 15.06 g/kg, total phosphorus mass ratio was 15.23 g/kg, total potassium mass ratio was 20.11 g/kg, mass ratio of alkaline hydrolysis nitrogen was 198.29 mg/kg, available phosphorus mass ratio was 36.22 mg/kg and available potassium mass ratio was 112.06 mg/kg.Figure 1Location of the study area. The map and inset map in this image were drawn by the authors using ArcGIS software. The software version used was ArcGIS software v.10.2, and its URL is http://www.esri.com/.Full size imageHumic acid fertilizerHumic acid fertilizer was produced by Yunnan Kunming Grey Environmental Protection Engineering Co., Ltd., China (Fig. 2). The organic matter was ≥ 61.4%, and the total nutrients (nitrogen, phosphorus and potassium) were ≥ 18.23%, of which N ≥ 3.63%, P2O5 ≥ 2.03%, and K2O ≥ 12.57%. The moisture content was ≤ 2.51%, the pH value was 5.7, the worm egg mortality rate was ≥ 95%, and the amount of faecal colibacillosis was ≤ 3%. The fertilizer contained numerous elements necessary for plants. The contents of harmful elements, including arsenic, mercury, lead, cadmium and chromium, were ≤ 2.8%, 0.01%, 7.6%, 0.1% and 4.7%, respectively; these were lower than the test standard.Figure 2Humic acid fertilizer in powder form.Full size imageExperimental design and observation methodsIrrigationIn this experiment, three irrigation practices, namely, control irrigation (C), wet irrigation (W) and flood irrigation (F), were designed (Table 1).Table 1 Different irrigation methods.Full size tableControl irrigation (C) of rice had no water layer in the rest of the growing stages, except for the shallow water layer at the regreen stage of rice, which was maintained at 0–30 mm, and the natural dryness in the yellow stage. The irrigation time and irrigation quota were determined by the root soil moisture content as the control index. The upper limit of irrigation was the saturated moisture content of the soil, the lower limit of soil moisture at each growth stage was the percentage of saturated moisture content, and the TPIME-PICO64/32 soil moisture analyser was used to determine the soil moisture content at 7:00 a.m. and 18:00 p.m., respectively. When the soil moisture content was close to or lower than the lower limit of irrigation, artificial irrigation occurred until the upper irrigation limit was reached. The soil moisture content was maintained between the upper irrigation limit and the lower irrigation limit of the corresponding fertility stage. Under the wet irrigation (W) and flood irrigation (F) conditions, it was necessary to read the depth of the water layer through bricks and a vertical ruler embedded in the field before and after 8:00 am every day to determine if irrigation was needed. If irrigation was needed, then the water metre was recorded before and after each irrigation. The difference between before and after was the amount of irrigation23.FertilizationIn our research, five fertilization methods were applied, as shown in Table 2. In this experiment, the rice cultivar “Suijing No. 18” was selected. Urea and humic acid fertilizer were applied according to the proportion of base fertilizer:tillering fertilizer:heading fertilizer (5:3:2). The amounts of phosphorus and potassium fertilizers were the same for all treatments, and P2O5 (45 kg ha−1) and K2O (80 kg ha−1) were used. Phosphorus was applied once as a basal application. Potassium fertilizer was applied twice: once as a basal fertilizer and at 8.5 leaf age (panicle primordium differentiation stage) at a 1:1 ratio22.Table 2 The fertilizer methods.Full size tableThis study was performed with a randomized complete block design with three replications. Three irrigation practices and five fertilizer methods were applied, for a total of 15 treatments as follows: CT1, CT2, CT3, CT4, CT5; WT1, WT2, WT3, WT4, WT5; FT1, FT2, FT3, FT4, and FT5 (C, W, and F represent control irrigation, wet irrigation, and flood irrigation; T represents fertilizer treatment).Measurements of the samplesA soil temperature sensor (HZTJ1-1) was buried in each experimental plot to monitor the temperature of each soil layer (5 cm, 10 cm, 15 cm, 20 cm and 25 cm depth). The transmission of photosynthetically active radiation was measured from 11:00 to 13:00 by using a SunScan Canopy Analysis System (Delta T Devices, Ltd., Cambridge, UK), and data during the crop-growing season were recorded every day24.Plant measurements were taken during the periods of tillering to ripening on days with no wind and good light. The fluorescence parameters were measured by a portable fluorescence measurement system (Li-6400XT, America). The detection light intensity was 1500 μmol m−2 s−1, and the saturated pulsed light intensity was 7200 μmolm−2 s−1. The functional leaves were dark adapted for 30 min, and then the maximum photosynthetic efficiency of PSII (Fv/Fm) was measured. Photochemical quenching (QP) and nonphotochemical quenching (NPQ) were measured with natural light. Simultaneously, the leaf chlorophyll relative content (SPAD) was monitored using SPAD 502 (Konica Minolta, Inc., Tokyo, Japan). For plant agronomic characteristics, the distance from the stem base to the stem tip was measured with a straight ruler to quantify plant height24.Statistical analysisExperimental data obtained for different parameters were analysed statistically using the analysis of variance technique as applicable to randomized complete block design. Duncan’s multiple range test was employed to assess differences between the treatment means at a 5% probability level. All statistical analyses were performed using SPSS 22.0 for Windows24.
    Ethics approvalExperimental research and field studies on plants, including the collection of plant material, comply with relevant institutional, national, and international guidelines and legislation. We had appropriate permissions/licences to perform the experiment in the study area. More