浙江农业学报 ›› 2025, Vol. 37 ›› Issue (9): 1881-1890.DOI: 10.3969/j.issn.1004-1524.20240872
收稿日期:2024-10-16
									
				
									
				
									
				
											出版日期:2025-09-25
									
				
											发布日期:2025-10-15
									
			作者简介:阎勇,E-mail:14544286@qq.com通讯作者:
					阎勇
							基金资助:
        
               		LUO Yanglan(
), HUANG Liling, HUANG Shilyu, YAN Yong(
), WANG Canqin
			  
			
			
			
                
        
    
Received:2024-10-16
									
				
									
				
									
				
											Online:2025-09-25
									
				
											Published:2025-10-15
									
			Contact:
					YAN Yong   
							摘要: 为优化羊肚菌(Morchella eximia)菌株WSJD-1的培养基配方并确定广西地区羊肚菌栽培关键技术,本研究采用Plackett-Burman设计、最陡爬坡试验和Box-Behnken响应面法优化菌株WSJD-1培养基。基于优化配方,探究不同播种期、播种量、覆膜方式和连作次数对广西地区羊肚菌生长的影响。结果表明:WSJD-1的最优培养基配方为小麦质量分数83.0%、木屑质量分数10.0%、谷壳质量分数1.8%、KH2PO4质量分数0.3%、MgSO4质量分数0.3%、腐殖土质量分数2.6%、生石灰质量分数1%、石膏质量分数1%,该配方下每袋菌核平均干重达20.879 g,较优化前提高14.68%。不同播种期、播种量、覆膜方式与连作次数对菌丝萌发、原基形成和幼菇出现时间影响均较小。广西地区羊肚菌最适播种期为11月25日至12月2日,每667 m2最佳播种量为200 kg。大棚外覆白色农膜+棚内覆黑色地膜处理的产量最高,达0.669 kg·m-2。连作导致产量显著下降,连作1、2、3次较未连作分别减产47.10%、56.38%和64.78%,且病虫害发生程度随连作次数增加而上升。本研究明确了WSJD-1菌株的最优培养基配方和广西地区羊肚菌栽培的关键技术参数,包括最适播种期、最佳播种量、最优覆膜方式与连作影响。
中图分类号:
罗阳兰, 黄丽玲, 黄世旅, 阎勇, 王灿琴. 羊肚菌WSJD-1的基质配方优化及其在广西地区的栽培技术[J]. 浙江农业学报, 2025, 37(9): 1881-1890.
LUO Yanglan, HUANG Liling, HUANG Shilyu, YAN Yong, WANG Canqin. Optimization of substrate formulation and cultivation techniques for Morchella eximia WSJD-1 in Guangxi region of China[J]. Acta Agriculturae Zhejiangensis, 2025, 37(9): 1881-1890.
| 水平 Level  |  各因素的质量分数Mass fraction of factors | |||||
|---|---|---|---|---|---|---|
| (A)小麦Wheat | (B)木屑Sawdust | (C)谷壳Chaff | (D)KH2PO4 | (E)MgSO4 | (F)腐殖土Muck | |
| -1 | 75 | 5 | 2.5 | 0.1 | 0.15 | 1.5 | 
| 1 | 85 | 10 | 5.0 | 0.2 | 0.30 | 2.5 | 
表1 Plackett-Burman试验设计
Table 1 Plackett-Burman experiments design %
| 水平 Level  |  各因素的质量分数Mass fraction of factors | |||||
|---|---|---|---|---|---|---|
| (A)小麦Wheat | (B)木屑Sawdust | (C)谷壳Chaff | (D)KH2PO4 | (E)MgSO4 | (F)腐殖土Muck | |
| -1 | 75 | 5 | 2.5 | 0.1 | 0.15 | 1.5 | 
| 1 | 85 | 10 | 5.0 | 0.2 | 0.30 | 2.5 | 
| 因素 Factor  |  各因素的质量分数Mass fraction of factors | ||
|---|---|---|---|
| (A)小麦 Wheat  |  (B)谷壳 Chaff  |  (C)KH2PO4 | |
| -1 | 75 | 1 | 0.20 | 
| 0 | 80 | 2 | 0.25 | 
| 1 | 85 | 3 | 0.30 | 
表2 Box-Behnken设计方案
Table 2 Box-Behnken design %
| 因素 Factor  |  各因素的质量分数Mass fraction of factors | ||
|---|---|---|---|
| (A)小麦 Wheat  |  (B)谷壳 Chaff  |  (C)KH2PO4 | |
| -1 | 75 | 1 | 0.20 | 
| 0 | 80 | 2 | 0.25 | 
| 1 | 85 | 3 | 0.30 | 
| 序号 Number  |  各因素的水平Level of factors | 每袋菌核干重 Dry weight of sclerotia per bag/g  | |||||
|---|---|---|---|---|---|---|---|
| A | B | C | D | E | F | ||
| 1 | -1 | 1 | 1 | -1 | 1 | -1 | 8.010 | 
| 2 | 1 | -1 | -1 | -1 | 1 | 1 | 18.045 | 
| 3 | -1 | 1 | -1 | -1 | -1 | 1 | 16.112 | 
| 4 | 1 | -1 | 1 | -1 | -1 | -1 | 11.254 | 
| 5 | -1 | -1 | -1 | 1 | 1 | 1 | 18.981 | 
| 6 | 1 | -1 | 1 | 1 | -1 | 1 | 17.012 | 
| 7 | 1 | 1 | 1 | -1 | 1 | 1 | 15.548 | 
| 8 | -1 | -1 | -1 | -1 | -1 | -1 | 9.784 | 
| 9 | -1 | -1 | 1 | 1 | 1 | -1 | 14.002 | 
| 10 | 1 | 1 | -1 | 1 | -1 | -1 | 18.855 | 
| 11 | 1 | 1 | -1 | 1 | 1 | -1 | 19.400 | 
| 12 | -1 | 1 | 1 | 1 | -1 | 1 | 11.447 | 
表3 Plackett-Burman设计试验结果
Table 3 Results of Plackett-Burman design
| 序号 Number  |  各因素的水平Level of factors | 每袋菌核干重 Dry weight of sclerotia per bag/g  | |||||
|---|---|---|---|---|---|---|---|
| A | B | C | D | E | F | ||
| 1 | -1 | 1 | 1 | -1 | 1 | -1 | 8.010 | 
| 2 | 1 | -1 | -1 | -1 | 1 | 1 | 18.045 | 
| 3 | -1 | 1 | -1 | -1 | -1 | 1 | 16.112 | 
| 4 | 1 | -1 | 1 | -1 | -1 | -1 | 11.254 | 
| 5 | -1 | -1 | -1 | 1 | 1 | 1 | 18.981 | 
| 6 | 1 | -1 | 1 | 1 | -1 | 1 | 17.012 | 
| 7 | 1 | 1 | 1 | -1 | 1 | 1 | 15.548 | 
| 8 | -1 | -1 | -1 | -1 | -1 | -1 | 9.784 | 
| 9 | -1 | -1 | 1 | 1 | 1 | -1 | 14.002 | 
| 10 | 1 | 1 | -1 | 1 | -1 | -1 | 18.855 | 
| 11 | 1 | 1 | -1 | 1 | 1 | -1 | 19.400 | 
| 12 | -1 | 1 | 1 | 1 | -1 | 1 | 11.447 | 
| 项 Term  |  效应 Effect  |  系数 Coefficient  |  系数标准误 Standard error of coefficient  |  t值 t value  |  p值 p value  | 
|---|---|---|---|---|---|
| 常量Constant | 14.871 | 0.521 | 28.55 | <0.001 | |
| A | 3.630 | 1.815 | 0.521 | 3.48 | 0.018 | 
| B | 0.049 | 0.024 | 0.521 | 0.05 | 0.964 | 
| C | -3.984 | -1.992 | 0.521 | -3.82 | 0.012 | 
| D | 3.491 | 1.745 | 0.521 | 3.35 | 0.020 | 
| E | 1.587 | 0.793 | 0.521 | 1.52 | 0.188 | 
| F | 2.640 | 1.320 | 0.521 | 2.53 | 0.052 | 
表4 方差分析结果
Table 4 Analysis of variance results
| 项 Term  |  效应 Effect  |  系数 Coefficient  |  系数标准误 Standard error of coefficient  |  t值 t value  |  p值 p value  | 
|---|---|---|---|---|---|
| 常量Constant | 14.871 | 0.521 | 28.55 | <0.001 | |
| A | 3.630 | 1.815 | 0.521 | 3.48 | 0.018 | 
| B | 0.049 | 0.024 | 0.521 | 0.05 | 0.964 | 
| C | -3.984 | -1.992 | 0.521 | -3.82 | 0.012 | 
| D | 3.491 | 1.745 | 0.521 | 3.35 | 0.020 | 
| E | 1.587 | 0.793 | 0.521 | 1.52 | 0.188 | 
| F | 2.640 | 1.320 | 0.521 | 2.53 | 0.052 | 
| 序号 Number  |  质量分数Mass fraction/% | 每袋菌核干重 Dry weight of sclerotia per bag/g  | ||
|---|---|---|---|---|
| 小麦 Wheat  |  谷壳 Chaff  |  KH2PO4 | ||
| 1 | 75 | 10 | 0.05 | 15.781 | 
| 2 | 77 | 8 | 0.10 | 16.946 | 
| 3 | 79 | 6 | 0.15 | 17.633 | 
| 4 | 81 | 4 | 0.20 | 18.309 | 
| 5 | 83 | 2 | 0.25 | 18.867 | 
| 6 | 85 | 1 | 0.30 | 17.801 | 
表5 最陡爬坡试验结果
Table 5 Results of the steepest ascent test
| 序号 Number  |  质量分数Mass fraction/% | 每袋菌核干重 Dry weight of sclerotia per bag/g  | ||
|---|---|---|---|---|
| 小麦 Wheat  |  谷壳 Chaff  |  KH2PO4 | ||
| 1 | 75 | 10 | 0.05 | 15.781 | 
| 2 | 77 | 8 | 0.10 | 16.946 | 
| 3 | 79 | 6 | 0.15 | 17.633 | 
| 4 | 81 | 4 | 0.20 | 18.309 | 
| 5 | 83 | 2 | 0.25 | 18.867 | 
| 6 | 85 | 1 | 0.30 | 17.801 | 
| 试验号 Test number  |  各因素的水平 Level of factors  |  每袋菌核干重 Dry weight of sclerotia per bag/g  | ||
|---|---|---|---|---|
| A | B | C | ||
| 1 | 0 | 0 | 0 | 19.993 | 
| 2 | 0 | 0 | 0 | 20.878 | 
| 3 | -1 | 0 | 1 | 16.895 | 
| 4 | 1 | 0 | -1 | 14.437 | 
| 5 | -1 | 1 | 0 | 16.279 | 
| 6 | 0 | -1 | -1 | 14.146 | 
| 7 | 0 | 0 | 0 | 21.001 | 
| 8 | 0 | 1 | -1 | 16.086 | 
| 9 | 0 | 1 | 1 | 16.988 | 
| 10 | 0 | 0 | 0 | 21.301 | 
| 11 | 1 | 0 | 1 | 19.660 | 
| 12 | 0 | 0 | 0 | 19.985 | 
| 13 | -1 | -1 | 0 | 15.001 | 
| 14 | 0 | -1 | 1 | 19.900 | 
| 15 | 1 | -1 | 0 | 17.259 | 
| 16 | 1 | 1 | 0 | 15.250 | 
| 17 | -1 | 0 | -1 | 13.768 | 
表6 响应面试验设计方案与结果
Table 6 Design and results of response surface experiment
| 试验号 Test number  |  各因素的水平 Level of factors  |  每袋菌核干重 Dry weight of sclerotia per bag/g  | ||
|---|---|---|---|---|
| A | B | C | ||
| 1 | 0 | 0 | 0 | 19.993 | 
| 2 | 0 | 0 | 0 | 20.878 | 
| 3 | -1 | 0 | 1 | 16.895 | 
| 4 | 1 | 0 | -1 | 14.437 | 
| 5 | -1 | 1 | 0 | 16.279 | 
| 6 | 0 | -1 | -1 | 14.146 | 
| 7 | 0 | 0 | 0 | 21.001 | 
| 8 | 0 | 1 | -1 | 16.086 | 
| 9 | 0 | 1 | 1 | 16.988 | 
| 10 | 0 | 0 | 0 | 21.301 | 
| 11 | 1 | 0 | 1 | 19.660 | 
| 12 | 0 | 0 | 0 | 19.985 | 
| 13 | -1 | -1 | 0 | 15.001 | 
| 14 | 0 | -1 | 1 | 19.900 | 
| 15 | 1 | -1 | 0 | 17.259 | 
| 16 | 1 | 1 | 0 | 15.250 | 
| 17 | -1 | 0 | -1 | 13.768 | 
| 方差来源 Source of variance  |  平方和 Sum of squares  |  自由度 Degree of freedom  |  均方 Mean square  |  F值 F value  |  p值 p value  | 
|---|---|---|---|---|---|
| 模型Model | 108.43 | 9 | 12.05 | 34.49 | < 0.000 1 | 
| A | 2.72 | 1 | 2.72 | 7.78 | 0.026 9 | 
| B | 0.36 | 1 | 0.36 | 1.04 | 0.342 2 | 
| C | 28.15 | 1 | 28.15 | 80.59 | < 0.000 1 | 
| AB | 2.70 | 1 | 2.70 | 7.73 | 0.027 3 | 
| AC | 1.10 | 1 | 1.10 | 3.14 | 0.119 5 | 
| BC | 5.89 | 1 | 5.89 | 16.85 | 0.004 5 | 
| A2 | 29.28 | 1 | 29.28 | 83.84 | < 0.000 1 | 
| B2 | 17.65 | 1 | 17.65 | 50.52 | 0.000 2 | 
| C2 | 13.71 | 1 | 13.71 | 39.25 | 0.000 4 | 
| 残差Residual | 2.44 | 7 | 0.35 | ||
| 失拟度 | 0.97 | 3 | 0.32 | 0.88 | 0.521 8 | 
| Lack of fit | |||||
| 绝对误差 | 1.47 | 4 | 0.37 | ||
| Pure error | |||||
| 总离差 | 110.87 | 16 | |||
| Total variation | 
表7 方差分析结果
Table 7 Results of variance analysis
| 方差来源 Source of variance  |  平方和 Sum of squares  |  自由度 Degree of freedom  |  均方 Mean square  |  F值 F value  |  p值 p value  | 
|---|---|---|---|---|---|
| 模型Model | 108.43 | 9 | 12.05 | 34.49 | < 0.000 1 | 
| A | 2.72 | 1 | 2.72 | 7.78 | 0.026 9 | 
| B | 0.36 | 1 | 0.36 | 1.04 | 0.342 2 | 
| C | 28.15 | 1 | 28.15 | 80.59 | < 0.000 1 | 
| AB | 2.70 | 1 | 2.70 | 7.73 | 0.027 3 | 
| AC | 1.10 | 1 | 1.10 | 3.14 | 0.119 5 | 
| BC | 5.89 | 1 | 5.89 | 16.85 | 0.004 5 | 
| A2 | 29.28 | 1 | 29.28 | 83.84 | < 0.000 1 | 
| B2 | 17.65 | 1 | 17.65 | 50.52 | 0.000 2 | 
| C2 | 13.71 | 1 | 13.71 | 39.25 | 0.000 4 | 
| 残差Residual | 2.44 | 7 | 0.35 | ||
| 失拟度 | 0.97 | 3 | 0.32 | 0.88 | 0.521 8 | 
| Lack of fit | |||||
| 绝对误差 | 1.47 | 4 | 0.37 | ||
| Pure error | |||||
| 总离差 | 110.87 | 16 | |||
| Total variation | 
																													图1 小麦、谷壳和KH2PO4质量分数对羊肚菌菌核干重影响的响应面结果 A,小麦质量分数;B谷壳质量分数;C, KH2PO4质量分数。
Fig.1 Response surface results of the effects of wheat, chaff, and KH2PO4 mass fraction on the dry weight of Morchella eximia sclerotia A, Wheat mass fraction; B, Chaff mass fraction; C, KH2PO4 mass fraction.
| 播种期 Sowing date  |  菌丝萌发时间 Mycelium germination time/d  |  原基形成时间 Primordium formation time/d  |  幼菇出现时间 Young fruiting body emergence time/d  |  生育期 Growth period/d  |  产量 Yield/(kg·m-2)  | 
|---|---|---|---|---|---|
| T1 | 4 | 45 | 54 | 65 | 0.383±0.019 e | 
| T2 | 5 | 44 | 53 | 66 | 0.485±0.017 d | 
| T3 | 5 | 45 | 56 | 66 | 0.584±0.034 c | 
| T4 | 7 | 47 | 55 | 68 | 0.703±0.030 a | 
| T5 | 7 | 48 | 56 | 68 | 0.716±0.012 a | 
| T6 | 9 | 47 | 56 | 67 | 0.661±0.071 b | 
| T7 | 9 | 46 | 57 | 67 | 0.436±0.048 d | 
| T8 | 12 | 44 | 56 | 68 | 0.423±0.039 d | 
表8 羊肚菌WSJD-1菌株在不同播种期的生长状况和产量
Table 8 Growth and yield of Morchella eximia WSJD-1 at different sowing dates
| 播种期 Sowing date  |  菌丝萌发时间 Mycelium germination time/d  |  原基形成时间 Primordium formation time/d  |  幼菇出现时间 Young fruiting body emergence time/d  |  生育期 Growth period/d  |  产量 Yield/(kg·m-2)  | 
|---|---|---|---|---|---|
| T1 | 4 | 45 | 54 | 65 | 0.383±0.019 e | 
| T2 | 5 | 44 | 53 | 66 | 0.485±0.017 d | 
| T3 | 5 | 45 | 56 | 66 | 0.584±0.034 c | 
| T4 | 7 | 47 | 55 | 68 | 0.703±0.030 a | 
| T5 | 7 | 48 | 56 | 68 | 0.716±0.012 a | 
| T6 | 9 | 47 | 56 | 67 | 0.661±0.071 b | 
| T7 | 9 | 46 | 57 | 67 | 0.436±0.048 d | 
| T8 | 12 | 44 | 56 | 68 | 0.423±0.039 d | 
| 播种量 Seeding rate  |  菌丝萌发时间 Mycelium germination time/d  |  原基形成时间 Primordium formation time/d  |  幼菇出现时间 Young fruiting body emergence time/d  |  生育期 Growth period/d  |  产量 Yield/(kg·m-2)  | 
|---|---|---|---|---|---|
| S1 | 7 | 47 | 56 | 67 | 0.689±0.016 a | 
| S2 | 6 | 47 | 55 | 66 | 0.677±0.023 a | 
| S3 | 7 | 49 | 59 | 68 | 0.514±0.018 b | 
表9 羊肚菌WSJD-1菌株在不同播种量下的生长状况和产量
Table 9 Growth and yield of Morchella eximia WSJD-1 under different seeding quantity treatments
| 播种量 Seeding rate  |  菌丝萌发时间 Mycelium germination time/d  |  原基形成时间 Primordium formation time/d  |  幼菇出现时间 Young fruiting body emergence time/d  |  生育期 Growth period/d  |  产量 Yield/(kg·m-2)  | 
|---|---|---|---|---|---|
| S1 | 7 | 47 | 56 | 67 | 0.689±0.016 a | 
| S2 | 6 | 47 | 55 | 66 | 0.677±0.023 a | 
| S3 | 7 | 49 | 59 | 68 | 0.514±0.018 b | 
| 覆膜方式 Mulching method  |  菌丝萌发时间 Mycelium germination time/d  |  原基形成时间 Primordium formation time/d  |  幼菇出现时间 Young fruiting body emergence time/d  |  生育期 Growth period/d  |  产量 Yield/(kg·m-2)  | 
|---|---|---|---|---|---|
| F1 | 6 | 45 | 54 | 67 | 0.418±0.025 b | 
| F2 | 7 | 44 | 53 | 66 | 0.431±0.019 b | 
| F3 | 7 | 47 | 55 | 68 | 0.301±0.016 c | 
| F4 | 6 | 46 | 56 | 67 | 0.669±0.027 a | 
表10 羊肚菌WSJD-1菌株在不同覆膜方式下的生长状况和产量
Table 10 Growth and yield of Morchella eximia WSJD-1 under different mulching methods
| 覆膜方式 Mulching method  |  菌丝萌发时间 Mycelium germination time/d  |  原基形成时间 Primordium formation time/d  |  幼菇出现时间 Young fruiting body emergence time/d  |  生育期 Growth period/d  |  产量 Yield/(kg·m-2)  | 
|---|---|---|---|---|---|
| F1 | 6 | 45 | 54 | 67 | 0.418±0.025 b | 
| F2 | 7 | 44 | 53 | 66 | 0.431±0.019 b | 
| F3 | 7 | 47 | 55 | 68 | 0.301±0.016 c | 
| F4 | 6 | 46 | 56 | 67 | 0.669±0.027 a | 
| 处理 Treatment  |  菌丝萌发时间 Mycelium germination time/d  |  原基形成时间 Primordium formation time/d  |  幼菇出现时间 Young fruiting body emergence time/d  |  生育期 Growth period/d  |  产量 Yield/(kg·m-2)  |  病虫害发生程度 Degree of occurrence of pests and diseases  | 
|---|---|---|---|---|---|---|
| R1 | 7 | 47 | 56 | 67 | 0.690±0.033 a | + | 
| R2 | 7 | 50 | 58 | 68 | 0.365±0.014 b | ++ | 
| R3 | 6 | 53 | 64 | 75 | 0.301±0.019 b | +++ | 
| R4 | 7 | 57 | 66 | 78 | 0.243±0.023 c | +++ | 
表11 不同连作次数处理下羊肚菌WSJD-1菌株的生长状况和产量
Table 11 Growth and yield of Morchella eximia WSJD-1 under different continuous cropping treatments
| 处理 Treatment  |  菌丝萌发时间 Mycelium germination time/d  |  原基形成时间 Primordium formation time/d  |  幼菇出现时间 Young fruiting body emergence time/d  |  生育期 Growth period/d  |  产量 Yield/(kg·m-2)  |  病虫害发生程度 Degree of occurrence of pests and diseases  | 
|---|---|---|---|---|---|---|
| R1 | 7 | 47 | 56 | 67 | 0.690±0.033 a | + | 
| R2 | 7 | 50 | 58 | 68 | 0.365±0.014 b | ++ | 
| R3 | 6 | 53 | 64 | 75 | 0.301±0.019 b | +++ | 
| R4 | 7 | 57 | 66 | 78 | 0.243±0.023 c | +++ | 
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