Epoxiconazole Improved Photosynthesis, Yield Formation, Grain Quality and 2-Acetyl-1-Pyrroline Biosynthesis of Fragrant Rice

doi:10.1016/j.rsci.2022.01.007

Abstract

Abstract:

Epoxiconazole is a triazole compound. However, the effects of epoxiconazole on crop productivity and quality were rarely reported. In this study, we investigated the effects of epoxiconazole application on yield formation, grain quality attributes, and 2-acetyl-1-pyrroline (2-AP) content in fragrant rice. A three-year field experiment was carried out with a fragrant rice variety, Meixiangzhan 2. At the heading stage, 0, 0.02, 0.04, 0.08, 0.16 and 0.32 g/L epoxiconazole solutions were foliar applied to fragrant rice plants, respectively. The results showed that epoxiconazole application significantly increased grain yield, seed-setting rate and 1000-grain weight. Chlorophyll content and net photosynthetic rate of fragrant rice during the grain-filling stage significantly increased due to epoxiconazole application. Foliar application of epoxiconazole at 0.08 g/L increased grain protein content and decreased both chalky rice rate and chalkiness area ratio of fragrant rice. Epoxiconazole also substantially increased grain 2-AP content by inducing the regulation in contents of related synthetic precursors, including proline, pyrroline-5- carboxylic acid, ∆1-pyrroline and methylglyoxal. Overall, foliar application of epoxiconazole could be used for the improvement in grain yield, grain quality and 2-AP content in fragrant rice production when applied concentration at 0.08-0.32 g/L. Our findings provided the new roles of epoxiconazole in crop production.

Key words: epoxiconazole, fragrant rice, 2-acetyl-1-pyrroline, yield formation, grain quality

Luo Haowen, He Longxin, Du Bin, Pan Shenggang, Mo Zhaowen, Yang Shuying, Zou Yingbin, Tang Xiangru. Epoxiconazole Improved Photosynthesis, Yield Formation, Grain Quality and 2-Acetyl-1-Pyrroline Biosynthesis of Fragrant Rice[J]. Rice Science, 2022, 29(2): 189-196.

Figures/Tables 6

References 32

[1]	Ajigboye O O, Murchie E, Ray R V. 2014. Foliar application of isopyrazam and epoxiconazole improves photosystem II efficiency, biomass and yield in winter wheat. Pest Biochem Physiol, 114: 52-60.
[2]	Alam M M, Tanaka T, Nakamura H, Ichikawa H, Kobayashi K, Yaeno T, Yamaoka N, Shimomoto K, Takayama K, Nishina H, Nishiguchi M. 2015. Overexpression of a rice heme activator protein gene (OsHAP2E) confers resistance to pathogens, salinity and drought, and increases photosynthesis and tiller number. Plant Biotechnol J, 13(1): 85-96.
[3]	Banu M N A, Hoque M A, Watanabe-Sugimoto M, Islam M M, Uraji M, Matsuoka K, Nakamura Y, Murata Y. 2010. Proline and glycinebetaine ameliorated NaCl stress via scavenging of hydrogen peroxide and methylglyoxal but not superoxide or nitric oxide in tobacco cultured cells. Biosci Biotechnol Biochem, 74(10): 2043-2049.
[4]	Bao G G, Ashraf U, Wang C L, He L X, Wei X S, Zheng A X, Mo Z W, Tang X R. 2018. Molecular basis for increased 2-acetyl- 1-pyrroline contents under alternate wetting and drying (AWD) conditions in fragrant rice. Plant Physiol Biochem, 133: 149-157.
[5]	Bertelsen J R, de Neergaard E, Smedegaard-Petersen V. 2001. Fungicidal effects of azoxystrobin and epoxiconazole on phyllosphere fungi, senescence and yield of winter wheat. Plant Pathol, 50(2): 190-205.
[6]	Chen Y, Yao J, Wang W X, Gao T C, Yang X, Zhang A F. 2013. Effect of epoxiconazole on rice blast and rice grain yield in China. Eur J Plant Pathol, 135(4): 675-682.
[7]	Duan M Y, Cheng S R, Lu R H, Lai R F, Zheng A X, Ashraf U, Fan P S, Du B, Luo H W, Tang X R. 2019. Effect of foliar sodium selenate on leaf senescence of fragrant rice in South China. Appl Ecol Environ Res, 17(2): 3343-3351.
[8]	Feng H Y, Jiang H L, Wang M, Tang X R, Duan M Y, Pan S G, Tian H, Wang S L, Mo Z W. 2019. Morphophysiological responses of different scented rice varieties to high temperature at seedling stage. Chin J Rice Sci, 33(1): 68-74. (in Chinese with English abstract)
[9]	Huang L C, Dai L P, Wang L, Leng Y J, Yang Y L, Xu J, Hu J, Rao Y C, Zhang G H, Zhu L, Dong G J, Guo L B, Qian Q, Zeng D L. 2015. Genetic dissection for chlorophyll content of the top three leaves during grain filling in rice (Oryza sativa L.). J Plant Growth Regul, 34(2): 381-391.
[10]	Kaziem A E, Gao B B, Li L S, Zhang Z X, He Z Z, Wen Y, Wang M H. 2020. Enantioselective bioactivity, toxicity, and degradation in different environmental mediums of chiral fungicide epoxiconazole. J Hazard Mater, 386: 121951. PMID
[11]	Kaziem A E, He Z Z, Li L S, Wen Y, Wang Z, Gao Y Y, Wang M H. 2021. Changes in soil and rat gut microbial diversity after long-term exposure to the chiral fungicide epoxiconazole. Chemosphere, 272: 129618. PMID
[12]	Kong L L, Ashraf U, Cheng S R, Rao G S, Mo Z W, Tian H, Pan S G, Tang X R. 2017. Short-term water management at early filling stage improves early-season rice performance under high temperature stress in South China. Eur J Agron, 90: 117-126.
[13]	Li M J, Ashraf U, Tian H, Mo Z W, Pan S G, Anjum S A, Duan M Y, Tang X R. 2016. Manganese-induced regulations in growth, yield formation, quality characters, rice aroma and enzyme involved in 2-acetyl-1-pyrroline biosynthesis in fragrant rice. Plant Physiol Biochem, 103: 167-175.
[14]	Liu S M, Fu L Y, Chen J P, Wang S, Liu J L, Jiang J, Che Z P, Tian Y E, Chen G Q. 2020. Baseline sensitivity and control efficacy of epoxiconazole against Fusarium graminearum in Henan Province, China. Eur J Plant Pathol, 157: 825-833.
[15]	Liu X W, Huang Z L, Li Y Z, Xie W J, Li W, Tang X R, Ashraf U, Kong L L, Wu L M, Wang S L, Mo Z W. 2020. Selenium- silicon (Se-Si) induced modulations in physio-biochemical responses, grain yield, quality, aroma formation and lodging in fragrant rice. Ecotoxicol Environ Saf, 196: 110525.
[16]	Luo H W, He L X, Du B, Wang Z M, Zheng A X, Lai R F, Tang X R. 2019. Foliar application of selenium (Se) at heading stage induces regulation of photosynthesis, yield formation, and quality characteristics in fragrant rice. Photosynthetica, 57(4): 1007-1014.
[17]	Luo H W, He L X, Du B, Pan S G, Mo Z W, Duan M Y, Tian H, Tang X R. 2020. Biofortification with chelating selenium in fragrant rice: Effects on photosynthetic rates, aroma, grain quality and yield formation. Field Crops Res, 255: 107909.
[18]	Mo Z W, Li W, Pan S G, Fitzgerald T L, Xiao F, Tang Y J, Wang Y L, Duan M Y, Tian H, Tang X R. 2015. Shading during the grain filling period increases 2-acetyl-1-pyrroline content in fragrant rice. Rice, 8: 9.
[19]	Mo Z W, Li Y H, Nie J, He L X, Pan S G, Duan M Y, Tian H, Xiao L Z, Zhong K Y, Tang X R. 2019. Nitrogen application and different water regimes at booting stage improved yield and 2-acetyl- 1-pyrroline (2AP) formation in fragrant rice. Rice, 12(1): 74.
[20]	Pan S G, Wen X C, Wang Z M, Ashraf U, Tian H, Duan M Y, Mo Z W, Fan P S, Tang X R. 2017. Benefits of mechanized deep placement of nitrogen fertilizer in direct-seeded rice in South China. Field Crops Res, 203: 139-149.
[21]	Pan Y Y, Chen Y B, Wang C R, Li H, Huang D Q, Zhou D G, Wang Z D, Zhao L, Gong R, Zhou S C. 2015. Metabolism of γ-aminobutyrate and 2-acetyl-1-pyrroline analyses at various grain developmental stages in rice (Oryza sativa L.). Chin J Rice Sci, 35(2): 121-129. (in Chinese with English abstract)
[22]	Poonlaphdecha J, Gantet P, Maraval I, Sauvage F X, Menut C, Morère A, Boulanger R, Wüst M, Gunata Z. 2016. Biosynthesis of 2-acetyl-1-pyrroline in rice calli cultures: Demonstration of 1-pyrroline as a limiting substrate. Food Chem, 197: 965-971. PMID
[23]	van den Bossche H, Willemsens G, Cools W, Marichal P, Lauwers W. 1983. Hypothesis on the molecular basis of the antifungal activity of N-substituted imidazoles and triazoles. Biochem Soc Trans, 11(6): 665-667.
[24]	Wakte K, Zanan R, Hinge V, Khandagale K, Nadaf A, Henry R. 2017. Thirty-three years of 2-acetyl-1-pyrroline, a principal basmati aroma compound in scented rice (Oryza sativa L.): A status review. J Sci Food Agric, 97(2): 384-395.
[25]	Wakte K V, Kad T D, Zanan R L, Nadaf A B. 2011. Mechanism of 2-acetyl-1-pyrroline biosynthesis in Bassia latifolia Roxb. flowers. Physiol Mol Biol Plants, 17(3): 231-237.
[26]	Xie W J, Kong L L, Ma L, Ashraf U, Pan S G, Duan M Y, Tian H, Wu L M, Tang X R, Mo Z W. 2020. Enhancement of 2-acetyl-1-pyrroline (2AP) concentration, total yield, and quality in fragrant rice through exogenous γ-aminobutyric acid (GABA) application. J Cereal Sci, 91: 102900.
[27]	Yan B P, Ye F, Gao D P. 2015. Residues of the fungicide epoxiconazole in rice and paddy in the Chinese field ecosystem. Pest Manag Sci, 71(1): 65-71.
[28]	Yoshihashi T, Huong N T T, Inatomi H. 2002. Precursors of 2-acetyl-1-pyrroline, a potent flavor compound of an aromatic rice variety. J Agric Food Chem, 50(7): 2001-2004.
[29]	Zhang W Y, Chen Y J, Wang Z Q, Yang J C. 2017. Polyamines and ethylene in rice young panicles in response to soil drought during panicle differentiation. Plant Growth Regul, 82: 491-503.
[30]	Zhang Z C, Zhang S F, Yang J C, Zhang J H. 2008. Yield, grain quality and water use efficiency of rice under non-flooded mulching cultivation. Field Crops Res, 108(1): 71-81.
[31]	Zhao R M, Luo H W, Wang Z M, Hu L. 2020. Benefits of continuous plow tillage to fragrant rice performance. Agron J, 112(5): 4171-4181.
[32]	Zhao Z X, Sun R X, Su Y, Hu J Y, Liu X L. 2021. Fate, residues and dietary risk assessment of the fungicides epoxiconazole and pyraclostrobin in wheat in twelve different regions, China. Ecotoxicol Environ Saf, 207: 111236.

Treatment	Grain yield (t/hm²)	No. of effective panicles per m²	No. of grains per panicle	Seed-setting rate (%)	1000-grain weight (g)
2018
CK	5.84 ± 0.11 bc	281.33 ± 9.07 a	147.88 ± 2.13 a	73.73 ± 1.73 c	19.84 ± 0.24 e
EP1	5.66 ± 0.14 c	278.00 ± 3.61 a	143.35 ± 1.88 a	73.55 ± 0.48 c	20.00 ± 0.41 de
EP2	5.97 ± 0.09 b	282.00 ± 11.53 a	146.65 ± 1.43 a	75.31 ± 1.37 bc	20.31 ± 0.25 cd
EP3	6.40 ± 0.08 a	284.00 ± 15.52 a	143.19 ± 2.63 a	77.91 ± 1.91 ab	20.46 ± 0.21 c
EP4	6.46 ± 0.09 a	279.67 ± 14.57 a	145.22 ± 2.29 a	80.22 ± 0.50 a	21.24 ± 0.27 a
EP5	6.41 ± 0.19 a	268.67 ± 13.28 a	140.57 ± 0.81 a	79.65 ± 1.75 a	20.85 ± 0.30 b
2019
CK	5.91 ± 0.08 c	267.33 ± 15.50 a	147.34 ± 2.16 a	74.57 ± 1.28 c	19.95 ± 0.16 d
EP1	5.73 ± 0.21 c	263.33 ± 14.05 a	144.96 ± 2.39 a	73.41 ± 0.96 c	19.64 ± 0.09 d
EP2	6.18 ± 0.16 b	276.33 ± 10.12 a	146.43 ± 2.68 a	74.53 ± 0.80 c	20.38 ± 0.24 c
EP3	6.38 ± 0.14 ab	268.33 ± 17.39 a	145.29 ± 2.53 a	78.10 ± 1.47 b	20.44 ± 0.20 bc
EP4	6.31 ± 0.17 ab	265.67 ± 20.23 a	147.17 ± 2.21 a	79.81 ± 0.72 a	21.19 ± 0.16 a
EP5	6.54 ± 0.09 a	293.00 ± 5.57 a	142.94 ± 2.37 a	80.42 ± 0.67 a	20.76 ± 0.21 b
2020
CK	5.77 ± 0.22 c	281.00 ± 15.62 a	147.81 ± 2.24 a	73.82 ± 1.41 c	19.99 ± 0.27 d
EP1	5.87 ± 0.10 bc	280.67 ± 19.86 a	148.66 ± 0.47 a	73.89 ± 1.48 c	20.27 ± 0.22 cd
EP2	6.10 ± 0.13 b	270.67 ± 16.74 a	141.31 ± 1.56 a	74.64 ± 0.90 c	20.44 ± 0.20 c
EP3	6.44 ± 0.14 a	283.67 ± 18.88 a	142.82 ± 1.83 a	77.69 ± 2.02 b	21.01 ± 0.03 b
EP4	6.52 ± 0.17 a	283.33 ± 17.01 a	144.78 ± 1.45 a	80.39 ± 0.50 a	21.13 ± 0.16 ab
EP5	6.50 ± 0.14 a	278.33 ± 18.34 a	145.81 ± 3.51 a	79.06 ± 0.83 ab	21.43 ± 0.32 a

Treatment	Grain yield (t/hm²)	No. of effective panicles per m²	No. of grains per panicle	Seed-setting rate (%)	1000-grain weight (g)
2018
CK	5.84 ± 0.11 bc	281.33 ± 9.07 a	147.88 ± 2.13 a	73.73 ± 1.73 c	19.84 ± 0.24 e
EP1	5.66 ± 0.14 c	278.00 ± 3.61 a	143.35 ± 1.88 a	73.55 ± 0.48 c	20.00 ± 0.41 de
EP2	5.97 ± 0.09 b	282.00 ± 11.53 a	146.65 ± 1.43 a	75.31 ± 1.37 bc	20.31 ± 0.25 cd
EP3	6.40 ± 0.08 a	284.00 ± 15.52 a	143.19 ± 2.63 a	77.91 ± 1.91 ab	20.46 ± 0.21 c
EP4	6.46 ± 0.09 a	279.67 ± 14.57 a	145.22 ± 2.29 a	80.22 ± 0.50 a	21.24 ± 0.27 a
EP5	6.41 ± 0.19 a	268.67 ± 13.28 a	140.57 ± 0.81 a	79.65 ± 1.75 a	20.85 ± 0.30 b
2019
CK	5.91 ± 0.08 c	267.33 ± 15.50 a	147.34 ± 2.16 a	74.57 ± 1.28 c	19.95 ± 0.16 d
EP1	5.73 ± 0.21 c	263.33 ± 14.05 a	144.96 ± 2.39 a	73.41 ± 0.96 c	19.64 ± 0.09 d
EP2	6.18 ± 0.16 b	276.33 ± 10.12 a	146.43 ± 2.68 a	74.53 ± 0.80 c	20.38 ± 0.24 c
EP3	6.38 ± 0.14 ab	268.33 ± 17.39 a	145.29 ± 2.53 a	78.10 ± 1.47 b	20.44 ± 0.20 bc
EP4	6.31 ± 0.17 ab	265.67 ± 20.23 a	147.17 ± 2.21 a	79.81 ± 0.72 a	21.19 ± 0.16 a
EP5	6.54 ± 0.09 a	293.00 ± 5.57 a	142.94 ± 2.37 a	80.42 ± 0.67 a	20.76 ± 0.21 b
2020
CK	5.77 ± 0.22 c	281.00 ± 15.62 a	147.81 ± 2.24 a	73.82 ± 1.41 c	19.99 ± 0.27 d
EP1	5.87 ± 0.10 bc	280.67 ± 19.86 a	148.66 ± 0.47 a	73.89 ± 1.48 c	20.27 ± 0.22 cd
EP2	6.10 ± 0.13 b	270.67 ± 16.74 a	141.31 ± 1.56 a	74.64 ± 0.90 c	20.44 ± 0.20 c
EP3	6.44 ± 0.14 a	283.67 ± 18.88 a	142.82 ± 1.83 a	77.69 ± 2.02 b	21.01 ± 0.03 b
EP4	6.52 ± 0.17 a	283.33 ± 17.01 a	144.78 ± 1.45 a	80.39 ± 0.50 a	21.13 ± 0.16 ab
EP5	6.50 ± 0.14 a	278.33 ± 18.34 a	145.81 ± 3.51 a	79.06 ± 0.83 ab	21.43 ± 0.32 a

Treatment	Proline content (μg/g)	P5C (μmol/g)	Δ1-pyrroline (μmol/g)	MG (μg/g)
2018
CK	11.18 ± 0.99 c	1.10 ± 0.02 b	0.94 ± 0.01 c	26.11 ± 1.36 a
EP1	11.07 ± 0.19 c	1.07 ± 0.06 b	0.91 ± 0.00 c	26.77 ± 1.02 a
EP2	12.94 ± 0.84 b	1.10 ± 0.02 b	0.93 ± 0.02 c	24.41 ± 1.66 a
EP3	16.12 ± 0.41 a	1.51 ± 0.15 a	1.05 ± 0.02 b	20.65 ± 1.35 b
EP4	16.39 ± 1.10 a	1.42 ± 0.15 a	1.16 ± 0.03 a	20.60 ± 0.11 b
EP5	16.51 ± 0.64 a	1.29 ± 0.15 a	1.17 ± 0.01 a	20.26 ± 0.68 b
2019
CK	11.06 ± 0.86 c	1.10 ± 0.04 b	0.92 ± 0.02 c	26.60 ± 1.38 a
EP1	10.91 ± 1.31 c	1.07 ± 0.06 b	0.97 ± 0.03 c	24.63 ± 0.35 b
EP2	13.49 ± 0.83 b	1.12 ± 0.01 b	0.94 ± 0.03 c	23.98 ± 0.19 b
EP3	16.41 ± 1.39 a	1.35 ± 0.16 a	1.07 ± 0.02 b	21.94 ± 0.71 c
EP4	16.68 ± 1.20 a	1.32 ± 0.17 a	1.14 ± 0.02 a	19.65 ± 1.67 d
EP5	17.43 ± 0.25 a	1.28 ± 0.16 a	1.12 ± 0.01 ab	19.66 ± 0.79 d
2020
CK	11.29 ± 0.72 c	1.05 ± 0.05 b	0.96 ± 0.03 c	26.97 ± 0.68 a
EP1	11.96 ± 1.07 bc	1.10 ± 0.04 b	0.98 ± 0.02 c	26.22 ± 1.53 a
EP2	13.00 ± 0.71 b	1.04 ± 0.04 b	0.95 ± 0.03 c	23.97 ± 1.69 a
EP3	16.81 ± 1.03 a	1.39 ± 0.17 a	1.06 ± 0.03 b	21.09 ± 1.32 b
EP4	15.29 ± 0.22 a	1.30 ± 0.23 a	1.17 ± 0.02 a	20.80 ± 1.16 b
EP5	16.46 ± 0.64 a	1.28 ± 0.07 a	1.15 ± 0.02 a	20.37 ± 1.84 b

Treatment	Proline content (μg/g)	P5C (μmol/g)	Δ1-pyrroline (μmol/g)	MG (μg/g)
2018
CK	11.18 ± 0.99 c	1.10 ± 0.02 b	0.94 ± 0.01 c	26.11 ± 1.36 a
EP1	11.07 ± 0.19 c	1.07 ± 0.06 b	0.91 ± 0.00 c	26.77 ± 1.02 a
EP2	12.94 ± 0.84 b	1.10 ± 0.02 b	0.93 ± 0.02 c	24.41 ± 1.66 a
EP3	16.12 ± 0.41 a	1.51 ± 0.15 a	1.05 ± 0.02 b	20.65 ± 1.35 b
EP4	16.39 ± 1.10 a	1.42 ± 0.15 a	1.16 ± 0.03 a	20.60 ± 0.11 b
EP5	16.51 ± 0.64 a	1.29 ± 0.15 a	1.17 ± 0.01 a	20.26 ± 0.68 b
2019
CK	11.06 ± 0.86 c	1.10 ± 0.04 b	0.92 ± 0.02 c	26.60 ± 1.38 a
EP1	10.91 ± 1.31 c	1.07 ± 0.06 b	0.97 ± 0.03 c	24.63 ± 0.35 b
EP2	13.49 ± 0.83 b	1.12 ± 0.01 b	0.94 ± 0.03 c	23.98 ± 0.19 b
EP3	16.41 ± 1.39 a	1.35 ± 0.16 a	1.07 ± 0.02 b	21.94 ± 0.71 c
EP4	16.68 ± 1.20 a	1.32 ± 0.17 a	1.14 ± 0.02 a	19.65 ± 1.67 d
EP5	17.43 ± 0.25 a	1.28 ± 0.16 a	1.12 ± 0.01 ab	19.66 ± 0.79 d
2020
CK	11.29 ± 0.72 c	1.05 ± 0.05 b	0.96 ± 0.03 c	26.97 ± 0.68 a
EP1	11.96 ± 1.07 bc	1.10 ± 0.04 b	0.98 ± 0.02 c	26.22 ± 1.53 a
EP2	13.00 ± 0.71 b	1.04 ± 0.04 b	0.95 ± 0.03 c	23.97 ± 1.69 a
EP3	16.81 ± 1.03 a	1.39 ± 0.17 a	1.06 ± 0.03 b	21.09 ± 1.32 b
EP4	15.29 ± 0.22 a	1.30 ± 0.23 a	1.17 ± 0.02 a	20.80 ± 1.16 b
EP5	16.46 ± 0.64 a	1.28 ± 0.07 a	1.15 ± 0.02 a	20.37 ± 1.84 b

Treatment	Brown rice rate (%)	Milled rice rate (%)	Head rice rate (%)	Protein content (%)	Amylose content (%)	Chalky rice rate (%)	Chalkiness area ratio (%)	Chalkiness degree
2018
CK	78.58 ± 0.34 a	66.00 ± 0.56 a	53.51 ± 0.33 a	8.63 ± 0.12 c	18.53 ± 0.15 c	10.45 ± 0.90 b	10.07 ± 0.40 ab	3.35 ± 0.61 a
EP1	79.23 ± 0.26 a	65.67 ± 1.01 a	53.43 ± 0.14 a	8.60 ± 0.10 c	18.57 ± 0.15 bc	10.83 ± 0.81 ab	9.80 ± 0.35 abc	2.54 ± 0.73 a
EP2	79.67 ± 0.14 a	66.13 ± 0.75 a	53.37 ± 0.13 a	8.87 ± 0.06 b	18.63 ± 0.12 bc	10.37 ± 0.60 b	8.41 ± 0.87 cd	2.70 ± 0.52 a
EP3	78.94 ± 0.65 a	66.31 ± 0.59 a	53.55 ± 0.17 a	9.10 ± 0.10 a	18.57 ± 0.12 bc	7.38 ± 0.99 c	7.90 ± 1.24 d	3.10 ± 0.39 a
EP4	78.84 ± 0.32 a	65.95 ± 0.66 a	53.71 ± 0.30 a	8.50 ± 0.10 c	18.73 ± 0.06 b	8.37 ± 0.74 c	9.15 ± 0.65 bcd	2.47 ± 0.29 a
EP5	78.77 ± 0.73 a	65.67 ± 0.47 a	53.27 ± 0.21 a	8.10 ± 0.10 d	19.20 ± 0.10 a	12.11 ± 0.78 a	10.92 ± 0.62 a	2.97 ± 0.68 a
2019
CK	78.67 ± 0.71 a	66.55 ± 0.13 a	53.50 ± 0.30 a	8.67 ± 0.06 b	18.43 ± 0.06 c	10.34 ± 1.12 b	10.92 ± 1.46 ab	3.34 ± 0.28 a
EP1	79.79 ± 0.05 a	66.17 ± 0.73 a	53.60 ± 0.49 a	8.63 ± 0.12 bc	18.53 ± 0.15 c	10.70 ± 0.93 b	9.85 ± 1.09 bc	2.84 ± 0.52 a
EP2	78.94 ± 0.40 a	66.45 ± 0.66 a	53.55 ± 0.10 a	8.80 ± 0.10 b	18.57 ± 0.15 c	8.38 ± 0.41 c	9.23 ± 0.88 bcd	2.96 ± 0.69 a
EP3	79.00 ± 0.82 a	66.08 ± 0.42 a	53.29 ± 0.28 a	9.20 ± 0.17 a	18.60 ± 0.17 c	6.35 ± 0.33 d	7.90 ± 0.60 d	2.74 ± 0.64 a
EP4	78.84 ± 0.37 a	66.03 ± 0.74 a	53.57 ± 0.42 a	8.43 ± 0.06 cd	18.90 ± 0.10 b	8.46 ± 1.30 c	9.15 ± 0.68 cd	2.45 ± 0.40 a
EP5	79.40 ± 0.61 a	65.78 ± 0.59 a	53.56 ± 0.13 a	8.23 ± 0.12 d	19.27 ± 0.06 a	12.18 ± 0.59 a	12.17 ± 0.28 a	2.94 ± 0.45 a
2020
CK	78.68 ± 0.57 a	65.80 ± 0.28 a	53.35 ± 0.30 a	8.60 ± 0.10 bc	18.57 ± 0.12 b	10.88 ± 0.58 ab	10.13 ± 1.07 ab	2.54 ± 0.41 b
EP1	79.11 ± 0.58 a	66.56 ± 0.14 a	53.54 ± 0.27 a	8.57 ± 0.12 bc	18.53 ± 0.06 b	10.03 ± 0.78 abc	11.52 ± 0.62 a	3.05 ± 0.26 ab
EP2	79.85 ± 0.11 a	65.71 ± 0.57 a	53.39 ± 0.16 a	8.80 ± 0.10 b	18.53 ± 0.15 b	9.43 ± 0.40 bcd	9.24 ± 1.33 bc	2.91 ± 0.67 ab
EP3	78.69 ± 0.13 a	65.70 ± 0.09 a	53.54 ± 0.20 a	9.10 ± 0.17 a	18.60 ± 0.10 b	7.92 ± 0.74 d	7.69 ± 1.26 c	3.00 ± 0.92 ab
EP4	79.01 ± 0.43 a	66.39 ± 0.42 a	53.41 ± 0.34 a	8.43 ± 0.15 c	18.70 ± 0.01 b	9.04 ± 1.51 cd	9.03 ± 0.81 bc	3.12 ± 0.15 ab
EP5	79.21 ± 0.54 a	65.67 ± 0.19 a	53.31 ± 0.08 a	8.10 ± 0.17 d	19.03 ± 0.06 a	11.07 ± 0.52 a	11.67 ± 0.87 a	3.78 ± 0.30 a