免耕和秸秆覆盖旱作稻田甲烷排放及甲烷产生菌群落结构和数量的变化特征
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摘要
水稻是我国南方主要的粮食作物。传统的淹水栽培水稻存在着两个重要的问题：水资源的大量消耗和甲烷的大量排放。因此,如何在保持水稻产量的同时减少水资源的消耗和甲烷的排放是研究者长期关注的热点问题。秸秆覆盖旱作可以有效减少水分的利用量,并可保持水稻的产量。然而,淹水条件下秸秆还田会显著增加甲烷排放,目前对秸秆覆盖旱作稻田甲烷排放规律的报道较少。免耕作为一种保护性耕作措施逐渐被广泛采用,最近有报道指出：免耕可以显著减少稻田甲烷的排放,但是对其中的机理研究尚在起步阶段。稻田甲烷排放是一个复杂的生理生化过程,甲烷产生菌是甲烷产生过程中必须的微生物类群,它在甲烷的产生和排放中起着重要作用,因此研究其群落结构和数量在水稻不同生育期及不同耕作模式下的变化特征可以更深入的了解甲烷的排放机理。
本试验利用2003年在江西省余江县建立的田间试验小区主要研究免耕和秸秆覆盖旱作稻田的甲烷排放规律及影响因素,并采用PCR-DGGE和荧光原位杂交(FISH)的方法对免耕及覆盖旱作稻田水稻不同生育期土壤中甲烷产生菌的群落结构和数量的变化特征进行研究,试图通过分析揭示甲烷产生菌群落结构和数量与稻田甲烷排放的关联性。
主要结果如下：
1、双季稻晚稻免耕稻田甲烷排放规律及其影响因子的研究
(1)在早稻生育期内,常耕处理的甲烷排放量为9.28g·m-2,免耕耕翻稻田的甲烷排放量为9.51g·m-2,二者之间没有显著差异；在晚稻生育期内,常耕处理的甲烷排放量为7.78g·m-2,免耕处理的甲烷排放量为3.04g·m-2,二者存在显著差异。这表明：免耕可以显著降低甲烷的排放量,免耕稻田耕翻后没有显著增加甲烷的排放。
(2)造成免耕稻田甲烷排放量减少的主要因素可能是免耕稻田表层土壤紧实度增加、土壤中可溶性有机碳含量降低、土壤甲烷产生菌数量减少和植株根系发育受到一定的限制等。
(3)在本研究中,晚稻的甲烷排放量低于早稻,与早稻相比常耕和免耕处理分别减少16%和68%。
2、秸秆覆盖旱作稻田的甲烷排放规律及覆盖旱作栽培模式的生产实践意义
(1)秸秆覆盖旱作、常规水作和无秸秆覆盖旱作稻田的在晚稻生育期内甲烷排放总量分别为：11.12g·m-2,7.78g·m-2和4.23g·m-2。与常规水作相比,覆盖旱作处理显著增加了甲烷排放,而无秸秆覆盖旱作稻田甲烷排放量显著减少。
(2)与传统水作相比,秸秆覆盖旱作可以显著降低水分的损失,有效提高水分的利用效率、显著降低表层土壤的温度。秸秆覆盖旱作可以显著提高土壤有机质、维持和提高土壤养分含量。秸秆覆盖旱作对水稻的产量没有明显影响,同时还可以大幅度提高水稻干物质累积量。因此,在水资源缺乏的地区,秸秆覆盖旱作是一种值得考虑的替代传统水作的水稻栽培模式。
3、不同管理措施稻田甲烷产生菌的群落结构和数量季节变化特征及其与甲烷排放的关系
Ⅰ、传统栽培模式双季稻稻田甲烷产生菌的群落结构和数量季节变化特征
(1)在传统栽培模式的双季稻稻田土壤中,甲烷产生菌的群落结构在早稻和晚稻生育期内均有一定的季节变化,其中有一部分主要的类群在整个生育期持续存在。早稻甲烷产生菌的丰富度,Shannon-Wiener多样性指数和均匀度分别为22,2.86和0.934,而晚稻的对应指标分别为19,2.66和0.916。
(2)在传统栽培模式的双季稻稻田土壤中,不同属甲烷产生菌的数量在水稻的不同生育期土壤中变化剧烈,不同时期的优势种也不相同。无论早稻还是晚稻,甲烷产生菌的总数均在水稻移栽20天左右达到最大,分别为1.15×107个/克干土和7.75×106个/克干土,并显著高于水稻的其它生长时期,其它生长时期甲烷产生菌的总数没有显著的差异。与早稻相比,晚稻甲烷产生菌的数量相对较少。
(3)无论是早稻还是晚稻,利用H2/CO2和甲酸盐的甲烷产生菌在营养类群上占优势,其次是利用醋酸盐的甲烷产生菌,再次是利用混合碳源的甲烷产生菌。Ⅱ、双季稻晚稻免耕处理稻田甲烷产生菌的群落结构和数量季节变化特征
(1)在双季稻晚稻免耕处理稻田中,早晚稻生育期内甲烷产生菌的群落结构也存在一定的季节变化,其中也有一部分主要的类群在整个生育期内持续存在；免耕处理早稻甲烷产生菌的丰富度,Shannon-Wiener多样性指数和均匀度分别为19,2.58和0.887,而晚稻的对应指标分别为21,2.78和0.884。
(2)在双季稻晚稻免耕处理稻田中,早稻和晚稻生育期内不同类群甲烷产生菌的数量也存在明显的季节变化。无论早稻还是晚稻,甲烷产生菌的总数也均在水稻移栽20天左右的分蘖旺盛期达到最大分别为1.14×107个/克干土和6.72×106个/克干。早稻分蘖期的甲烷产生菌数量显著高于其它采样时间的数量。而晚稻分蘖期甲烷产生菌数量仅显著高于晚稻移栽前和收获时,与其它两个生育期在数量上没有显著性差异。同样其它采样时期其数量没有显著差异。这种单季免耕的双季稻晚稻的甲烷产生菌数量也少于早稻。
(3)与传统栽培模式相比,单季免耕模式双季稻早稻甲烷产生菌群落结构和数量均偏低,而晚稻生长季土壤中甲烷产生菌的群落结构多样性则较高,但是数量仍低于常耕处理。
(4)免耕处理的早稻和晚稻的甲烷产生菌主要营养类型也为利用H2/CO2和甲酸盐的类群,这类甲烷产生菌显著多于其它营养类型,而利用醋酸盐和混合碳源的甲烷产生菌较少,而这二者之间没有显著性的差别。Ⅲ、秸秆覆盖旱作稻田甲烷产生茵的群落结构和数量季节变化特征
(1)覆盖旱作稻田和无覆盖旱作稻田甲烷产生菌的群落结构也有一定的季节变化,其中也有一部分主要的类群在整个生育期持续存在。覆盖旱作稻田甲烷产生菌的群落结构比无秸秆覆盖旱作稻田的多样性,丰富度高。
(2)覆盖旱作稻田和无覆盖旱作稻田甲烷产生菌的总数也均在水稻移栽后21天时达到最大,此时秸秆覆盖旱作稻田的甲烷产生菌数量显著高于无秸秆覆盖旱作稻田,而其它时期两个处理之间没有显著性差异。
(3)无论是秸秆覆盖旱作稻田还是无秸秆覆盖旱作稻田,在营养类群上利用H2/CO2和甲酸盐的甲烷产生菌数量显著高于其它营养类型的数量,其次是利用混合碳源的甲烷产生菌,再次是利用醋酸盐的甲烷产生菌。
(4)与常规淹水稻田相比,秸秆覆盖旱作稻田和无秸秆覆盖旱作稻田的甲烷产生菌的群落结构多样性和数量均偏小。但是三个处理甲烷产生菌的数量均在水稻移栽后21天达到最大,分别为7.75×106个/克干土、3.45×106个/克干土和4.21×106个/克干土。而群落结构Shannon-Wiener多样性指数的最大值出现的时间却有明显的差异。Ⅳ稻田甲烷排放与甲烷产生菌群落结构和数量的关系
稻田甲烷排放速率与稻田土壤甲烷产生菌的数量存在显著的正相关关系,而与土壤中甲烷产生菌群落结构的丰富度和Shannon-Wiener多样性指数的相关性较小
Rice (Oryza sativa L) is the main foodstuffs in south China. However, conventional rice cultivation consumes plentiful of water and flooded paddy field is a main methane emission source. It is important to seek an alternative rice cultivation style which could keep high rice yield but lower water consumption and methane emission. Thus, many experiments were conducted to investigate the relationship between the water management and rice yield and try to explore new rice cultivation style which could keep high rice yield but low water consumption in rice cultivation. The results suggested that non-flooded with straw mulching could reduce the water consumption, and keep high crop yield or even improve the crop yield. Unfortunately, straw application could increase CH4emission largely in paddy field in previous studies. However, mid-season drainage or intermittent drainage management in paddy field could reduce the methane emission significantly. So the methane emission from non-flooded with straw mulching paddy field may much less than the flooded with straw mulching paddy field. However, there was no information about the methane emission from non-flooded with straw mulching paddy field.
No-till is widely used as conservation tillage practice in dry land farming systems, but there were fewer reports in paddy field. The recent studies showed that no-till could reduce methane emission significantly, but the mechanism was not clearly.
Methanogens, which belong to the Euruarchaeota in archaeal, catalyze the last step in the anoxic degradation of organic matter for the formation of CH4. So, it is important to reveal the ecology of methanogenic archaeal in paddy field soil.
Thus, a water management field experiment was conducted in Yujiang County, Jiangxi Province in the southeast of China (N28°15, E116°55). The treatments included conventional flooded cultivation (F), non-flooded cultivation without straw mulching (NF-ZM) and with straw mulching (NF-M). We investigated the methane emission pattern in non-flooded with and without rice straw mulching paddy field. We also discussed the integrated effects of non-flooded with straw mulching rice cultivation style by investigating the rice yield, biomass accumulation, water use efficiency and soil nutrient balance.
A tillage practice field experiment with late rice no-till was conducted in a rice-rice crop system. We investigated the methane emission pattern in a double-rice cropping system under the contrasting tillage systems. Meanwhile, we investigated the soil organic carbon and dissolved organic carbon content and soil bulk density in order to validate whether the dissolved organic carbon content reduced under no-tillage as speculated in previous studies.
We also studied the methanogens community structure by PCR-DGGE and the population dynamic of specific species methanogens by fluorescence in situ hybridization (FISH) in different rice growing period under double rice cropping system and different field managements'paddy field.
The main results were followed:
First, the methane emission pattern and mechanisms in double rice cropping system under conventional and low tillage
1. The seasonal amount methane emission in the early rice growing period of T-T and T-NT was9.28g-m-2and9.51g·m-2, respectively. The T-NT didn't increase methane emission significantly after tillage in no-tillage field. The maximum emission rate of T-T and T-NT was18.52mg·m-2·h-1and7.32mg·m-2·h-1, respectively in late rice. The methane emission rate of T-NT was significant lower than T-T in the late rice growing period analysis by paired-sample T test. The seasonal amount methane emission of T-T and T-NT was7.78g·m-2and3.04g·m-2, respectively in late rice. The T-NT leads to a61%reduction of the total methane emission compared to the T-T treatment in late rice.
2. The no-till increased the surface soil organic matter content, but not significantly. But the increased soil organic matter reduced with the tillage and rice growth. No-till reduced soil active organic carbon content, especially the dissolved organic carbon (DOC). No-till significantly also increased the surface soil (0-10cm) bulk density. Meanwhile, the root parameters under no-till were lower than conventional tillage, and the differences were significant in last half rice growing period. The lower methane emission flux under no-till may due to the increased soil bulk density, lower dissolved organic carbon content, the lower number of methanogens and the restriction of root growth.
3. Compared to early rice growing period, the seasonal amount methane emission in late rice of T-T and T-NT reduced16%and68%, respectively. This may be due to the five months'fallow after last season late rice and long time rain before early rice transplanting.
Second, the methane emission pattern in non-flooded with straw mulching paddy field and the perspective of non-flooded with straw mulching rice cultivation
1. The peak methane flux appeared approximately2-3weeks after transplanting in all the treatments. The seasonal amount methane emission of the conventional flooded cultivation (F), non-flooded cultivation without straw mulching (NF-ZM) and with straw mulching (NF-M) was7.78g·m-2,4.23g·m-2and11.12g·m-2, respectively. In comparison with F, NF-M increased seasonal CH4emissions by43%, but NF-ZM reduced seasonal CH4emissions by46%. For the non-flooded treatments, the seasonal amount methane emissions of NF-M were2.62times higher than that of NF-ZM in this study.
2. The water content of10cm depth soil of NF-M was significant higher than that of NF-ZM in this study. Straw mulching can debase the surface soil temperature, which can reduce the damage of hot temperature on plant growth in summer. In comparison with F, the NF-M increased the soil organic matter content, soil total N and available K content significantly. The soil AH-N and Olsen-P of NF-M were also higher than F, but not significantly. All the soil nutrients investigated in this study of NF-M were significant higher than that of NF-ZM. These indicated that straw mulching could maintain and improve soil quality.
3. The yield of the NF-M, F and NF-ZM was9.60t·ha-1,9.75t-ha'and8.15t·ha-1respectively. This indicated that straw mulching could improve rice growth and maintain rice yield under non-flooded condition. The straw mulching could mitigate the water stress under non-flooded paddy field, there were no significant differences in rice growth character compared to F. The biomasses of NF-M were significantly higher than NF-ZM and F.
4. The straw mulching non-flooded cultivation increased the methane emission, but this cultivation style could get an almost equivalent yield with the conventional flooded cultivation style besides reducing water consumption significantly, improving soil quality and maintaining soil productivity. Furthermore, it avoided the greenhouse gas emission and atmosphere pollution during the straw management by other methods. Though non-flooded without straw mulching reduced methane emission significantly, it also reduced rice yield significantly. Thus, non-flooded with straw mulching rice cultivation style may be an alternative cultivation style in water shortage area.
Third, The dynamic of the community structure and population of methanogens under different field management paddy fields
I The dynamic of the community structure and population of methanogens in the double rice cropping system of south China
1. It is proper to use primer1106F-gc-1378R for PCR amplification of16S rRNA gene fragments of methanogenic archaeal for community structure analysis. The denaturant gradient range of the gel was from32to62%. The fluorescence in situ hybridization (FISH) is a feasible method to investigate the population of methanogens.
2. The community structure of methanogens changed greatly in different rice growing period in both early and late rice. There were several main species in all the growing period. The methanogens richness, diversity index and evenness of early rice were21.8,2.86and0.934respectively, and the parameters of late rice were18.5,2.66and0.916respectively. The community diversity of methanogens in early rice was higher than late rice.
3. The number of different methanogens species changed greatly in different rice growing period, the key methanogens varied in different rice growing period. The peak number of methanogens of early rice was1.15×10/In/g dry soil, and the lowest number was6.28×106In/g dry soil. The peak number of methanogens of late rice was7.75×106In/g dry soil. The peak number all appeared20days after transplanting. The total number of methanogens reached highest in tillering period in both early and late rice, and significantly higher than other rice growing period. There was no significantly difference in different rice growing period except tillering period.
4. The key physiological group of methanogens genera was using H2/CO2and formate, the followed was using acetate and then mixture substrate.
II the dynamic of the community structure and population of methanogens in a double rice cropping system under conventional and low tillage
1. The community structure of methanogens changed greatly in different rice growing period under no-till in both early and late rice. There were several main species in all the growing period under no-till. The methanogens richness, diversity index and evenness under no-till in early rice were19,2.58and0.887respectively, and the parameters of late rice were21,2.78and0.884respectively. These indicated that the community structure diversity of early rice was lower than that of late rice under no-till.
2. The population of different methanogens also changed greatly in different rice growing period under no-till in both early and late rice. In early rice, the total number of methanogens increased after rice transplanting and reached highest20days after rice transplanting at the density of1.14×107Ind·g-1dry soil. Thereafter, the number decreased slowly and reached lowest at the harvest time at the density of4.93×106Ind·g-1dry soil. In late rice, the the total number of methanogens also increased after rice transplanting and reached highest20days after rice transplanting at the density of6.72×106Ind·g-1dry soil. The total number of methanogens in early rice was higher than that of late rice. The peak number all appeared20days after transplanting as conventional tillage paddy field. There were no significant differences between the different rice growing periods except tillering period (about20days after transplanting).
3. The community strcture diversity and population under no-till was lower than conventional tillage paddy field in early rice. The community strcture diversity under no-till was higher than that of conventional tillage paddy field in late rice, but the number was also lower than conventional tillage paddy field.
4. The key physiological group of methanogens genera was using H2/CO2and formate under no-till in both early and late rice. The followed was using acetate and mixture substrate, and there was no significant difference between them.
HI the dynamic of the community structure and population of methanogens under non-flooded with and without straw mulching paddy field
1. The community structure of methanogens changed greatly in different rice growing period under non-flooded with straw mulching or without straw mulching. There were also several main species in all the growing period under non-flooded condition. The methanogens richness, diversity index and evenness under NF-M were25,2.89and0.950respectively, and the parameters under NF-ZM were21,2.79and0.916respectively. These indicated that the community structure diversity under NF-ZM was lower than that of NF-M.
2. The significant difference of the number of methanogens under NF-ZM and NF-M only appeared21days after transplanting. The total number of methanogens under NF-M was significantly higher than that under NF-ZM according the whole rice growing season.
3. In comparison with conventional flooded paddy field, the community structure diverisity and population under non-flooded with or without straw mulching were much lower. But the peak number of the three treatments all appeared21days after rice transplanting. The maximum values of diversity index of non-flooded fields were significantly late than conventional flooded one.
4. The key physiological group of methanogens genera was using H2/CO2and formate under both of NF-ZM and NF-M, the followed was using mixture substrate and then acetate.
IV the relationship between methane emission and the community structure and population of methanogens in paddy field
The methane emission rate was significant positive correation with the number of methanogens under different field managements' paddy fields, but the methane emission rate was negative correlation with the community structure of methanogens. This indicated that methane emission rate was mainly affected by the number of methanogens rather than the community structure of methanogens in paddy field.
本试验利用2003年在江西省余江县建立的田间试验小区主要研究免耕和秸秆覆盖旱作稻田的甲烷排放规律及影响因素,并采用PCR-DGGE和荧光原位杂交(FISH)的方法对免耕及覆盖旱作稻田水稻不同生育期土壤中甲烷产生菌的群落结构和数量的变化特征进行研究,试图通过分析揭示甲烷产生菌群落结构和数量与稻田甲烷排放的关联性。
主要结果如下：
1、双季稻晚稻免耕稻田甲烷排放规律及其影响因子的研究
(1)在早稻生育期内,常耕处理的甲烷排放量为9.28g·m-2,免耕耕翻稻田的甲烷排放量为9.51g·m-2,二者之间没有显著差异；在晚稻生育期内,常耕处理的甲烷排放量为7.78g·m-2,免耕处理的甲烷排放量为3.04g·m-2,二者存在显著差异。这表明：免耕可以显著降低甲烷的排放量,免耕稻田耕翻后没有显著增加甲烷的排放。
(2)造成免耕稻田甲烷排放量减少的主要因素可能是免耕稻田表层土壤紧实度增加、土壤中可溶性有机碳含量降低、土壤甲烷产生菌数量减少和植株根系发育受到一定的限制等。
(3)在本研究中,晚稻的甲烷排放量低于早稻,与早稻相比常耕和免耕处理分别减少16%和68%。
2、秸秆覆盖旱作稻田的甲烷排放规律及覆盖旱作栽培模式的生产实践意义
(1)秸秆覆盖旱作、常规水作和无秸秆覆盖旱作稻田的在晚稻生育期内甲烷排放总量分别为：11.12g·m-2,7.78g·m-2和4.23g·m-2。与常规水作相比,覆盖旱作处理显著增加了甲烷排放,而无秸秆覆盖旱作稻田甲烷排放量显著减少。
(2)与传统水作相比,秸秆覆盖旱作可以显著降低水分的损失,有效提高水分的利用效率、显著降低表层土壤的温度。秸秆覆盖旱作可以显著提高土壤有机质、维持和提高土壤养分含量。秸秆覆盖旱作对水稻的产量没有明显影响,同时还可以大幅度提高水稻干物质累积量。因此,在水资源缺乏的地区,秸秆覆盖旱作是一种值得考虑的替代传统水作的水稻栽培模式。
3、不同管理措施稻田甲烷产生菌的群落结构和数量季节变化特征及其与甲烷排放的关系
Ⅰ、传统栽培模式双季稻稻田甲烷产生菌的群落结构和数量季节变化特征
(1)在传统栽培模式的双季稻稻田土壤中,甲烷产生菌的群落结构在早稻和晚稻生育期内均有一定的季节变化,其中有一部分主要的类群在整个生育期持续存在。早稻甲烷产生菌的丰富度,Shannon-Wiener多样性指数和均匀度分别为22,2.86和0.934,而晚稻的对应指标分别为19,2.66和0.916。
(2)在传统栽培模式的双季稻稻田土壤中,不同属甲烷产生菌的数量在水稻的不同生育期土壤中变化剧烈,不同时期的优势种也不相同。无论早稻还是晚稻,甲烷产生菌的总数均在水稻移栽20天左右达到最大,分别为1.15×107个/克干土和7.75×106个/克干土,并显著高于水稻的其它生长时期,其它生长时期甲烷产生菌的总数没有显著的差异。与早稻相比,晚稻甲烷产生菌的数量相对较少。
(3)无论是早稻还是晚稻,利用H2/CO2和甲酸盐的甲烷产生菌在营养类群上占优势,其次是利用醋酸盐的甲烷产生菌,再次是利用混合碳源的甲烷产生菌。Ⅱ、双季稻晚稻免耕处理稻田甲烷产生菌的群落结构和数量季节变化特征
(1)在双季稻晚稻免耕处理稻田中,早晚稻生育期内甲烷产生菌的群落结构也存在一定的季节变化,其中也有一部分主要的类群在整个生育期内持续存在；免耕处理早稻甲烷产生菌的丰富度,Shannon-Wiener多样性指数和均匀度分别为19,2.58和0.887,而晚稻的对应指标分别为21,2.78和0.884。
(2)在双季稻晚稻免耕处理稻田中,早稻和晚稻生育期内不同类群甲烷产生菌的数量也存在明显的季节变化。无论早稻还是晚稻,甲烷产生菌的总数也均在水稻移栽20天左右的分蘖旺盛期达到最大分别为1.14×107个/克干土和6.72×106个/克干。早稻分蘖期的甲烷产生菌数量显著高于其它采样时间的数量。而晚稻分蘖期甲烷产生菌数量仅显著高于晚稻移栽前和收获时,与其它两个生育期在数量上没有显著性差异。同样其它采样时期其数量没有显著差异。这种单季免耕的双季稻晚稻的甲烷产生菌数量也少于早稻。
(3)与传统栽培模式相比,单季免耕模式双季稻早稻甲烷产生菌群落结构和数量均偏低,而晚稻生长季土壤中甲烷产生菌的群落结构多样性则较高,但是数量仍低于常耕处理。
(4)免耕处理的早稻和晚稻的甲烷产生菌主要营养类型也为利用H2/CO2和甲酸盐的类群,这类甲烷产生菌显著多于其它营养类型,而利用醋酸盐和混合碳源的甲烷产生菌较少,而这二者之间没有显著性的差别。Ⅲ、秸秆覆盖旱作稻田甲烷产生茵的群落结构和数量季节变化特征
(1)覆盖旱作稻田和无覆盖旱作稻田甲烷产生菌的群落结构也有一定的季节变化,其中也有一部分主要的类群在整个生育期持续存在。覆盖旱作稻田甲烷产生菌的群落结构比无秸秆覆盖旱作稻田的多样性,丰富度高。
(2)覆盖旱作稻田和无覆盖旱作稻田甲烷产生菌的总数也均在水稻移栽后21天时达到最大,此时秸秆覆盖旱作稻田的甲烷产生菌数量显著高于无秸秆覆盖旱作稻田,而其它时期两个处理之间没有显著性差异。
(3)无论是秸秆覆盖旱作稻田还是无秸秆覆盖旱作稻田,在营养类群上利用H2/CO2和甲酸盐的甲烷产生菌数量显著高于其它营养类型的数量,其次是利用混合碳源的甲烷产生菌,再次是利用醋酸盐的甲烷产生菌。
(4)与常规淹水稻田相比,秸秆覆盖旱作稻田和无秸秆覆盖旱作稻田的甲烷产生菌的群落结构多样性和数量均偏小。但是三个处理甲烷产生菌的数量均在水稻移栽后21天达到最大,分别为7.75×106个/克干土、3.45×106个/克干土和4.21×106个/克干土。而群落结构Shannon-Wiener多样性指数的最大值出现的时间却有明显的差异。Ⅳ稻田甲烷排放与甲烷产生菌群落结构和数量的关系
稻田甲烷排放速率与稻田土壤甲烷产生菌的数量存在显著的正相关关系,而与土壤中甲烷产生菌群落结构的丰富度和Shannon-Wiener多样性指数的相关性较小
Rice (Oryza sativa L) is the main foodstuffs in south China. However, conventional rice cultivation consumes plentiful of water and flooded paddy field is a main methane emission source. It is important to seek an alternative rice cultivation style which could keep high rice yield but lower water consumption and methane emission. Thus, many experiments were conducted to investigate the relationship between the water management and rice yield and try to explore new rice cultivation style which could keep high rice yield but low water consumption in rice cultivation. The results suggested that non-flooded with straw mulching could reduce the water consumption, and keep high crop yield or even improve the crop yield. Unfortunately, straw application could increase CH4emission largely in paddy field in previous studies. However, mid-season drainage or intermittent drainage management in paddy field could reduce the methane emission significantly. So the methane emission from non-flooded with straw mulching paddy field may much less than the flooded with straw mulching paddy field. However, there was no information about the methane emission from non-flooded with straw mulching paddy field.
No-till is widely used as conservation tillage practice in dry land farming systems, but there were fewer reports in paddy field. The recent studies showed that no-till could reduce methane emission significantly, but the mechanism was not clearly.
Methanogens, which belong to the Euruarchaeota in archaeal, catalyze the last step in the anoxic degradation of organic matter for the formation of CH4. So, it is important to reveal the ecology of methanogenic archaeal in paddy field soil.
Thus, a water management field experiment was conducted in Yujiang County, Jiangxi Province in the southeast of China (N28°15, E116°55). The treatments included conventional flooded cultivation (F), non-flooded cultivation without straw mulching (NF-ZM) and with straw mulching (NF-M). We investigated the methane emission pattern in non-flooded with and without rice straw mulching paddy field. We also discussed the integrated effects of non-flooded with straw mulching rice cultivation style by investigating the rice yield, biomass accumulation, water use efficiency and soil nutrient balance.
A tillage practice field experiment with late rice no-till was conducted in a rice-rice crop system. We investigated the methane emission pattern in a double-rice cropping system under the contrasting tillage systems. Meanwhile, we investigated the soil organic carbon and dissolved organic carbon content and soil bulk density in order to validate whether the dissolved organic carbon content reduced under no-tillage as speculated in previous studies.
We also studied the methanogens community structure by PCR-DGGE and the population dynamic of specific species methanogens by fluorescence in situ hybridization (FISH) in different rice growing period under double rice cropping system and different field managements'paddy field.
The main results were followed:
First, the methane emission pattern and mechanisms in double rice cropping system under conventional and low tillage
1. The seasonal amount methane emission in the early rice growing period of T-T and T-NT was9.28g-m-2and9.51g·m-2, respectively. The T-NT didn't increase methane emission significantly after tillage in no-tillage field. The maximum emission rate of T-T and T-NT was18.52mg·m-2·h-1and7.32mg·m-2·h-1, respectively in late rice. The methane emission rate of T-NT was significant lower than T-T in the late rice growing period analysis by paired-sample T test. The seasonal amount methane emission of T-T and T-NT was7.78g·m-2and3.04g·m-2, respectively in late rice. The T-NT leads to a61%reduction of the total methane emission compared to the T-T treatment in late rice.
2. The no-till increased the surface soil organic matter content, but not significantly. But the increased soil organic matter reduced with the tillage and rice growth. No-till reduced soil active organic carbon content, especially the dissolved organic carbon (DOC). No-till significantly also increased the surface soil (0-10cm) bulk density. Meanwhile, the root parameters under no-till were lower than conventional tillage, and the differences were significant in last half rice growing period. The lower methane emission flux under no-till may due to the increased soil bulk density, lower dissolved organic carbon content, the lower number of methanogens and the restriction of root growth.
3. Compared to early rice growing period, the seasonal amount methane emission in late rice of T-T and T-NT reduced16%and68%, respectively. This may be due to the five months'fallow after last season late rice and long time rain before early rice transplanting.
Second, the methane emission pattern in non-flooded with straw mulching paddy field and the perspective of non-flooded with straw mulching rice cultivation
1. The peak methane flux appeared approximately2-3weeks after transplanting in all the treatments. The seasonal amount methane emission of the conventional flooded cultivation (F), non-flooded cultivation without straw mulching (NF-ZM) and with straw mulching (NF-M) was7.78g·m-2,4.23g·m-2and11.12g·m-2, respectively. In comparison with F, NF-M increased seasonal CH4emissions by43%, but NF-ZM reduced seasonal CH4emissions by46%. For the non-flooded treatments, the seasonal amount methane emissions of NF-M were2.62times higher than that of NF-ZM in this study.
2. The water content of10cm depth soil of NF-M was significant higher than that of NF-ZM in this study. Straw mulching can debase the surface soil temperature, which can reduce the damage of hot temperature on plant growth in summer. In comparison with F, the NF-M increased the soil organic matter content, soil total N and available K content significantly. The soil AH-N and Olsen-P of NF-M were also higher than F, but not significantly. All the soil nutrients investigated in this study of NF-M were significant higher than that of NF-ZM. These indicated that straw mulching could maintain and improve soil quality.
3. The yield of the NF-M, F and NF-ZM was9.60t·ha-1,9.75t-ha'and8.15t·ha-1respectively. This indicated that straw mulching could improve rice growth and maintain rice yield under non-flooded condition. The straw mulching could mitigate the water stress under non-flooded paddy field, there were no significant differences in rice growth character compared to F. The biomasses of NF-M were significantly higher than NF-ZM and F.
4. The straw mulching non-flooded cultivation increased the methane emission, but this cultivation style could get an almost equivalent yield with the conventional flooded cultivation style besides reducing water consumption significantly, improving soil quality and maintaining soil productivity. Furthermore, it avoided the greenhouse gas emission and atmosphere pollution during the straw management by other methods. Though non-flooded without straw mulching reduced methane emission significantly, it also reduced rice yield significantly. Thus, non-flooded with straw mulching rice cultivation style may be an alternative cultivation style in water shortage area.
Third, The dynamic of the community structure and population of methanogens under different field management paddy fields
I The dynamic of the community structure and population of methanogens in the double rice cropping system of south China
1. It is proper to use primer1106F-gc-1378R for PCR amplification of16S rRNA gene fragments of methanogenic archaeal for community structure analysis. The denaturant gradient range of the gel was from32to62%. The fluorescence in situ hybridization (FISH) is a feasible method to investigate the population of methanogens.
2. The community structure of methanogens changed greatly in different rice growing period in both early and late rice. There were several main species in all the growing period. The methanogens richness, diversity index and evenness of early rice were21.8,2.86and0.934respectively, and the parameters of late rice were18.5,2.66and0.916respectively. The community diversity of methanogens in early rice was higher than late rice.
3. The number of different methanogens species changed greatly in different rice growing period, the key methanogens varied in different rice growing period. The peak number of methanogens of early rice was1.15×10/In/g dry soil, and the lowest number was6.28×106In/g dry soil. The peak number of methanogens of late rice was7.75×106In/g dry soil. The peak number all appeared20days after transplanting. The total number of methanogens reached highest in tillering period in both early and late rice, and significantly higher than other rice growing period. There was no significantly difference in different rice growing period except tillering period.
4. The key physiological group of methanogens genera was using H2/CO2and formate, the followed was using acetate and then mixture substrate.
II the dynamic of the community structure and population of methanogens in a double rice cropping system under conventional and low tillage
1. The community structure of methanogens changed greatly in different rice growing period under no-till in both early and late rice. There were several main species in all the growing period under no-till. The methanogens richness, diversity index and evenness under no-till in early rice were19,2.58and0.887respectively, and the parameters of late rice were21,2.78and0.884respectively. These indicated that the community structure diversity of early rice was lower than that of late rice under no-till.
2. The population of different methanogens also changed greatly in different rice growing period under no-till in both early and late rice. In early rice, the total number of methanogens increased after rice transplanting and reached highest20days after rice transplanting at the density of1.14×107Ind·g-1dry soil. Thereafter, the number decreased slowly and reached lowest at the harvest time at the density of4.93×106Ind·g-1dry soil. In late rice, the the total number of methanogens also increased after rice transplanting and reached highest20days after rice transplanting at the density of6.72×106Ind·g-1dry soil. The total number of methanogens in early rice was higher than that of late rice. The peak number all appeared20days after transplanting as conventional tillage paddy field. There were no significant differences between the different rice growing periods except tillering period (about20days after transplanting).
3. The community strcture diversity and population under no-till was lower than conventional tillage paddy field in early rice. The community strcture diversity under no-till was higher than that of conventional tillage paddy field in late rice, but the number was also lower than conventional tillage paddy field.
4. The key physiological group of methanogens genera was using H2/CO2and formate under no-till in both early and late rice. The followed was using acetate and mixture substrate, and there was no significant difference between them.
HI the dynamic of the community structure and population of methanogens under non-flooded with and without straw mulching paddy field
1. The community structure of methanogens changed greatly in different rice growing period under non-flooded with straw mulching or without straw mulching. There were also several main species in all the growing period under non-flooded condition. The methanogens richness, diversity index and evenness under NF-M were25,2.89and0.950respectively, and the parameters under NF-ZM were21,2.79and0.916respectively. These indicated that the community structure diversity under NF-ZM was lower than that of NF-M.
2. The significant difference of the number of methanogens under NF-ZM and NF-M only appeared21days after transplanting. The total number of methanogens under NF-M was significantly higher than that under NF-ZM according the whole rice growing season.
3. In comparison with conventional flooded paddy field, the community structure diverisity and population under non-flooded with or without straw mulching were much lower. But the peak number of the three treatments all appeared21days after rice transplanting. The maximum values of diversity index of non-flooded fields were significantly late than conventional flooded one.
4. The key physiological group of methanogens genera was using H2/CO2and formate under both of NF-ZM and NF-M, the followed was using mixture substrate and then acetate.
IV the relationship between methane emission and the community structure and population of methanogens in paddy field
The methane emission rate was significant positive correation with the number of methanogens under different field managements' paddy fields, but the methane emission rate was negative correlation with the community structure of methanogens. This indicated that methane emission rate was mainly affected by the number of methanogens rather than the community structure of methanogens in paddy field.
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