利用DGGE法研究不同种植体系中根际微生物群落结构

利用DGGE技术研究不同间作和轮作种植体系对作物根际细菌和真菌群落结构的影响.运用16SrDNA和18SrDNA特异引物对,将土壤中提取的总DNA进行PCR扩增后,通过DGGE技术对PCR产物进行分析,结果表明:玉米-蚕豆轮作对蚕豆根际细菌和真菌群落结构影响明显,二者都与单作蚕豆有较大差异;小麦/蚕豆间作明显改变两种作物根际细菌群落结构和蚕豆根际真菌群落结构;玉米/蚕豆间作明显改变玉米根际细菌、真菌群落结构和蚕豆根际真菌群落结构.     

小麦蚕豆间作对蚕豆根际微生物群落功能多样性的影响及其与蚕豆枯萎病发生的关系

通过田间小区试验,研究了小麦与蚕豆间作对蚕豆枯萎病发生和根际微生物代谢功能多样性的影响。结果表明,小麦与蚕豆间作使蚕豆枯萎病的发病率和病情指数分别比单作显著降低20%和30.4%。与单作处理相比,间作显著增加了蚕豆和小麦根际微生物对31种碳源的平均利用率（AWCD）,其中间作蚕豆的AWCD值最高,比单作增加82.7%,单作蚕豆最低。间作蚕豆和间作小麦根际微生物的Shannon多样性指数与丰富度指数均显著高于单作,间作使蚕豆和小麦的丰富度指数分别增加29.2%和30.3%。根际微生物对六类碳源的利用强度百分比以糖类、羧酸类和氨基酸最高,分别为41.96%,19.80%和18.13%。主成分分析表明,小麦与蚕豆间作改变了根际微生物的群落组成;相关分析表明,糖类、羧酸类和氨基酸类碳源是区分单间作处理差异的主要碳源,其中氨基酸类碳源是最敏感的碳源。小麦与蚕豆间作增加了根际微生物活性,提高了Shannon多样性指数和丰富度指数,改变了微生物群落功能多样性,是抑制蚕豆枯萎病的有效措施。该研究为阐明根际微生物功能多样性变化在间作体系病害控制中的作用与机制奠定了理论基础。     

不同品种小麦与蚕豆间作对蚕豆枯萎病的防治及其机理

通过田间小区试验和水培试验,研究了3个品种小麦（云麦42、云麦47和绵阳29）与蚕豆间作对蚕豆地上部生物量、枯萎病发生、根际微生物代谢功能多样性和枯萎病病原菌尖孢镰刀菌数量的影响,分析了3个不同小麦品种低分子量根系分泌物组分（糖、氨基酸和有机酸）的差异.结果表明： 云麦42与蚕豆间作（YM42/B）和云麦47与蚕豆间作（YM47/B）使蚕豆地上部生物量分别增加16.6%和13.4%,使蚕豆枯萎病病情指数分别降低47.6%和23.3%,绵阳29与蚕豆间作（MY29/B）对蚕豆地上部生物量和枯萎病病情指数均无显著影响.与单作蚕豆相比,YM42/B和YM47/B处理均显著提高了蚕豆根际微生物的平均颜色变化率（AWCD)及对碳源的总利用强度和Biolog ECO板中各种碳源的利用,明显改变了蚕豆根际微生物群落结构,并显著降低了蚕豆根际的尖孢镰刀菌数量;MY29/B处理对AWCD值、碳源总利用强度和Biolog ECO板中各种碳源的利用及蚕豆根际尖孢镰刀菌数量均无显著影响,也未明显改变蚕豆根际的微生物群落结构.3个小麦品种根系分泌物中可溶性糖、游离氨基酸总量和有机酸总量均表现为MY29＞YM47＞YM42.MY29根系分泌物中丝氨酸、谷氨酸、甘氨酸、缬氨酸、蛋氨酸、苯丙氨酸、赖氨酸的含量均显著高于YM42和YM47;精氨酸为YM42、YM47所特有,而亮氨酸为MY29所特有.MY29和YM47根系分泌物中检出酒石酸、苹果酸、柠檬酸、琥珀酸、延胡索酸和顺乌头酸6种有机酸,而YM42根系分泌物中检出洒石酸、苹果酸、柠檬酸和延胡索酸4种有机酸,且MY29和YM47根系分泌物中苹果酸含量显著高于YM42.小麦与蚕豆间作提高了蚕豆根际微生物的活性和碳源利用强度,改变了根际微生物的群落结构,降低了蚕豆根际尖孢镰刀菌的数量,促进了蚕豆生长,抑制了蚕豆枯萎病的发生,但小麦与蚕豆间作控病效果受小麦品种的影响,表明间作系统中非寄主作物根系分泌物的差异是影响间作对土传病害控制的重要因素.     

间作缓解蚕豆连作障碍的根际微生态效应

通过田间小区试验,研究了3个品种蚕豆(92-24、云豆324、凤豆6号)与小麦间作对蚕豆产量、枯萎病病情指数、根际镰刀菌数量、根际真菌代谢功能多样性和土壤酶活性的影响。结果表明:与单作蚕豆相比,云豆324与小麦间作(YD324/W)和凤豆6号与小麦间作(FD6/W)处理均显著提高了蚕豆地上部干重、籽粒产量和百粒重。YD324/W和FD6/W处理使蚕豆枯萎病发病初期病情指数分别降低57.14%和41.67%,镰刀菌数量分别降低32.06%和29.88%,而92-24与小麦间作(92-24/W)处理蚕豆产量、枯萎病病情指数和镰刀菌数量与单作蚕豆均无显著差异。YD324/W和FD6/W处理显著提高了蚕豆根际真菌的多样性指数和丰富度指数,并使蚕豆根际真菌的AWCD值分别比单作蚕豆提高了61.75%和46.49%;YD324/W和FD6/W处理明显改变了蚕豆根际真菌的群落结构。而92-24/W处理对蚕豆根际真菌的多样性指数、丰富度指数和AWCD值均无显著影响,也未明显改变真菌的群落结构。不同发病时期,YD324/W和FD6/W处理均显著提高了蚕豆根际土壤的蔗糖酶、脲酶和过氧化氢酶活性;而92-24/W处理蚕豆根际蔗糖酶、脲酶和过氧化氢酶活性与单作蚕豆均无显著差异。总之,小麦与不同品种蚕豆间作改变了蚕豆根际的真菌群落结构,提高了蚕豆根际真菌的活性、多样性和丰富度,提高土壤酶活性并改善蚕豆生长,增加了蚕豆产量。表明小麦与蚕豆间作改善了根际土壤的微生态环境,降低了镰刀菌的数量,缓解了蚕豆连作障碍,但蚕豆品种的差异影响间作控病效果。     

小麦-蚕豆间作条件下氮肥施用量对根际微生物区系的影响

通过田间小区试验,研究了小麦/蚕豆间作条件下4种施氮水平(0、90、180和270 kg·hm^-2)对根际微生物区系和多样性的影响.结果表明：在整个生育期,微生物数量有一定的波动,但均以开花期数量最高.与单作相比,间作显著增加了小麦和蚕豆根际的细菌、真菌、放线菌数量和微生物总量,而显著降低了开花期和成熟期蚕豆根际的微生物多样性.在不施氮（N₀）和低氮（N₉₀）水平下,间作与单作在微生物数量上的差异较大.间作对土壤微生物数量的促进效应在分蘖期和开花期最大,成熟期显著降低.小麦根际的微生物数量随施氮量的增加先增加后降低,以常规施氮处理（N₁₈₀）的微生物数量最多,氮肥用量对单作小麦的影响大于间作小麦. 施氮量对蚕豆根际细菌、真菌、放线菌数量和微生物多样性无显著影响,但降低了间作蚕豆根际微生物总量.适量施用氮肥能有效调节根际微生物区系,间作系统地上部植物多样性与地下部微生物区系间存在紧密联系.     

间作减轻蚕豆枯萎病的微生物和生理机制

通过田间小区试验,研究了小麦与蚕豆间作对蚕豆枯萎病发病率、病情指数、根际镰刀菌数量、蚕豆根系抗氧化酶活性和膜质过氧化的影响.采用Biolog ECO板分析了根际土壤微生物的代谢功能多样性,通过高效液相色谱(HPLC)测定了蚕豆根际土壤中酚酸含量.结果表明: 与单作蚕豆相比,小麦与蚕豆间作有降低蚕豆枯萎病发病率的趋势;发病盛期和发病末期,间作使蚕豆枯萎病的病情指数比单作显著降低15.8%和22.8%,明显提高了蚕豆根际微生物活性(AWCD值),根际微生物的Shannon多样性指数显著提高4.4%和5.3%,丰富度指数显著提高19.4%和37.1%.主成分分析表明,发病盛期和发病末期,间作明显改变了蚕豆根际微生物的群落结构,蚕豆根际镰刀菌数量分别降低53.8%和33.1%;并显著降低了蚕豆根际土壤中对羟基苯甲酸、香草酸、丁香酸、阿魏酸、苯甲酸和肉桂酸的含量.发病盛期和发病末期,间作蚕豆根系的过氧化物酶(POD)和过氧化氢酶(CAT)活性分别显著提高了20.0%、31.3%和38.5%、66.7%,丙二醛(MDA)含量显著降低36.3%和46.3%;发病初期间作对蚕豆根系的POD、CAT酶活性和MDA含量无显著影响.小麦与蚕豆间作显著提高了蚕豆根际微生物的活性、多样性和根系抗氧化酶活性,降低了蚕豆根际土壤中酚酸含量和膜质过氧化程度,减少了土壤镰刀菌的数量,从而提高了蚕豆对枯萎病的抗性,降低了枯萎病的危害程度.     

间作栽培对连作马铃薯根际土壤微生物群落的影响

连作严重影响了作物的产量和品质,而土壤微生物群落结构与功能对土壤生态系统和植物健康至关重要。以连作10a土壤为基质,单作马铃薯为对照,采用磷脂脂肪酸(Phospholipid fatty acids)、BIOLOGA技术和真菌形态学鉴定方法,研究了玉米、蚕豆与马铃薯间作模式下土壤微生物群落结构、功能和丛植菌根(Arbuscular Mycorrhizal)真菌对土壤环境变化的响应。结果表明:间作调控下,马铃薯根际土壤微生物主要类群结构发生显著改变;玉米间作马铃薯,土壤微生物群落总生物量降低,但群落功能多样性提高,促进了以羧酸类、多聚化合物、芳香类化合物、氨基酸类化合物为碳源的微生物类群代谢活性增强;蚕豆间作增加了土壤微生物总生物量,仅促进了以碳水化合物为碳源的微生物类群代谢活性。间作改变了作物根际土壤AM真菌的种、属数,AM真菌多样性降低,优势种由明球囊霉、地球囊霉转变为玉米间作体系里的福摩萨球囊霉、球泡球囊霉;蚕豆间作体系里福摩萨球囊霉和疣状无梗囊霉是优势种。间作栽培下AM真菌优势种群的变化可能受植物间的共生关系、微生物结构与功能等因素的制约。间作条件下,玉米显著影响了马铃薯根际土壤微生物群落功能多样性,而蚕豆则显著改变了微生物群落结构多样性;玉米、蚕豆对马铃薯根际土壤微生物群落功能与结构变化的影响不同步;间作调控后持续的土壤微生物群落结构与功能观察才有助于解释土壤微生物结构变化引起的功能响应。     

玉米、小麦和大麦与蚕豆间作体系不同根系分隔方式对蚕豆结瘤的影响

本实验选取3种对土壤氮素竞争能力不同的禾本科作物大麦(Hordeum vulgare)、小麦(Triticum aestivum)和玉米(Zea mays)分别与蚕豆(Vicia faba)间作, 建立对土壤氮素竞争能力不同的作物组合; 并采用3种分隔方式(塑料膜分隔、尼龙网分隔和无分隔)建立同一作物组合条件下作物种间根系相互作用的不同强度, 来研究不同作物组合及种间根系相互作用强度对蚕豆结瘤的影响。结果如下: (1)蚕豆的结瘤并未随3种禾本科作物氮素竞争能力的增强而增加, 但是3种间作体系蚕豆的结瘤却均表现出无分隔处理多于塑料膜分隔处理, 即同一间作体系种间根系相互作用越强, 越有利于蚕豆结瘤的产生, 存在种间互利作用; (2)在玉米/蚕豆间作体系中, 无分隔处理的蚕豆根瘤数目和根瘤重显著高于塑料膜分隔处理, 分别高出67.5%和70.1%; 在大麦/蚕豆间作体系中也表现出无分隔处理的根瘤重显著高于塑料膜分隔处理(高出46.3%); (3)玉米/蚕豆间作体系与小麦/蚕豆和大麦/蚕豆间作体系相比, 无分隔处理时土壤氮素含量显著高于后2个间作体系, 但是玉米/蚕豆间作体系对蚕豆结瘤的促进作用更强。上述结果表明, 在蚕豆/玉米间作体系中, 玉米促进蚕豆生物固氮除了氮素竞争机制外, 还可能存在其它机制。     

玉米、小麦和大麦与蚕豆间作体系不同根系分隔方式对蚕豆结瘤的影响

本实验选取3种对土壤氮素竞争能力不同的禾本科作物大麦(Hordeum vulgare)、小麦(Triticum aestivum)和玉米(Zea mays)分别与蚕豆(Vicia faba)间作,建立对土壤氮素竞争能力不同的作物组合;并采用3种分隔方式(塑料膜分隔、尼龙网分隔和无分隔)建立同一作物组合条件下作物种间根系相互作用的不同强度,来研究不同作物组合及种间根系相互作用强度对蚕豆结瘤的影响。结果如下:(1)蚕豆的结瘤并未随3种禾本科作物氮素竞争能力的增强而增加,但是3种间作体系蚕豆的结瘤却均表现出无分隔处理多于塑料膜分隔处理,即同一间作体系种间根系相互作用越强,越有利于蚕豆结瘤的产生,存在种间互利作用;(2)在玉米/蚕豆间作体系中,无分隔处理的蚕豆根瘤数目和根瘤重显著高于塑料膜分隔处理,分别高出67.5%和70.1%;在大麦/蚕豆间作体系中也表现出无分隔处理的根瘤重显著高于塑料膜分隔处理(高出46.3%);(3)玉米/蚕豆间作体系与小麦/蚕豆和大麦/蚕豆间作体系相比,无分隔处理时土壤氮素含量显著高于后2个间作体系,但是玉米/蚕豆间作体系对蚕豆结瘤的促进作用更强。上述结果表明,在蚕豆/玉米间作体系中,玉米促进蚕豆生物固氮除了氮素竞争机制外,还可能存在其它机制。     

间作大豆对甘蔗根际土壤细菌及固氮菌多样性的影响

为探讨间作大豆(Glycine max)对甘蔗(Saccharum officinarum)根际土壤细菌及固氮细菌多样性的影响, 收集和开发固氮菌资源, 筛选高效甘蔗联合固氮体系, 选用3个甘蔗栽培品种‘ROC22’、‘GT21’、‘B8’与大豆品种‘Guizao 2’进行间种栽培, 采用巢式PCR特异扩增细菌16S rRNA基因片段和固氮细菌nifH基因片段, 并结合变性梯度凝胶电泳(DGGE)技术, 对间作大豆的甘蔗根际土壤细菌及固氮细菌进行系统演化和多样性分析。聚类分析结果显示, 间作大豆改变了甘蔗根际土壤细菌及固氮细菌原来的群落组成结构, 尤其对固氮菌群落组成的改变更大, 但对群落物种的优势度影响较小。Shannon-Wiener多样性指数和Simpson多样性指数分析结果表明, 甘蔗-大豆间作显著影响甘蔗根际土壤中细菌和固氮菌的多样性, 其中对固氮细菌多样性的影响较大。不同甘蔗品种的根际土壤细菌和固氮菌在间作大豆条件下表现出不同的多样性, ‘ROC22’和‘GT21’间作处理甘蔗根际土壤固氮细菌的Shannon-Wiener多样性指数显著高于单作处理, 而‘ROC22’与大豆间作处理的甘蔗根际土壤固氮菌多样性最为丰富。在大豆生长盛期, 间作处理的甘蔗根际土壤细菌多样性最为丰富, 不同处理间的差异也最大, 随后下降。总体来看, 甘蔗-大豆间作显著地影响根际土壤细菌和固氮菌的群落结构和群落多样性, 有助于对甘蔗合理间作栽培模式的认识和筛选高效甘蔗联合固氮体系。     

Nitrogen Fixation of Faba Bean (Vicia faba L.) Interacting with a Non-legume in Two Contrasting Intercropping Systems

A field experiment was carried out to quantify biological nitrogen fixation (BNF) using the ¹⁵N isotope natural abundance method in maize (Zea mays L.)/faba bean (Vicia faba L.) and wheat (Triticum aestivum L.)/faba bean intercropping systems. Faba bean was yielding more in the maize/faba bean intercropping, but not in the wheat/faba bean intercropping. Biomass, grain yield and N acquisition of faba bean were significantly increased when intercropped with maize, and decreased significantly with wheat, irrespective of N-fertilizer application, indicating that the legume could gain or lose productivity in an intercropping situation. There was yield advantage of maize/faba bean intercropping, but no in wheat/faba bean intercropping. The grain yield of the faba bean intercropped with maize was greater than that of faba bean monoculture due to increases of the stems per plant and the pods per stem of faba bean. N fertilization inhibited N fixation of faba bean in maize/faba bean and wheat/faba bean intercropping and faba bean monoculture. The responses of different cropping systems to N-fertilizer application, however, were not identical, with competitive intercropping (wheat/faba bean) being more sensitive than facilitative intercropping (maize/faba bean). Intercropping increased the percentage of N derived from air (%Ndfa) of the wheat/faba bean system, but not that of the maize/faba bean system when no N fertilizer was applied. When receiving 120 kg N/ha, however, intercropping did not significantly increase %Ndfa either in the wheat/faba bean system or in the maize/faba bean system in comparison with faba bean in monoculture. The amount of shoot N derived from air (Ndfa), however, increased significantly when intercropped with maize, irrespective of N-fertilizer application. Ndfa decreased when intercropped with wheat, albeit not significantly at 120 kg N/ha. Ndfa was correlated more closely with dry matter yield, grain yield and competitive ratio, than with %Ndfa. This indicates that that total dry matter yield (sink strength), not %Ndfa, was more critical for the legume to increase Ndfa. The results suggested that N fixation could be improved by yield maximization in an intercropping system.     

Using competitive and facilitative interactions in intercropping systems enhances crop productivity and nutrient-use efficiency

This paper reviews recent research on the processes involved in the yield advantage in wheat (Triticum aestivum L.)/maize (Zea mays L.), wheat/soybean [Glycine max (L.) Merr.], faba bean (Vicia faba L.)/maize, peanut (Arachis hypogaea L.)/maize and water convolvulus (Ipomoea aquatica Forsk.)/maize intercropping. In wheat/maize and wheat/soybean intercropping systems, a significant yield increase of intercropped wheat over sole wheat was observed, which resulted from positive effects of the border row and inner rows of intercropped wheat. The border row effect was due to interspecific competition for nutrients as wheat had a higher competitive ability than either maize or soybean had. There was also compensatory growth, or a recovery process, of subordinate species such as maize and soybean, offsetting the impairment of early growth of the subordinate species. Finally, both dominant and subordinate species in intercropping obtain higher yields than that in corresponding sole wheat, maize or soybean. We summarized these processes as the `competition-recovery production principle'. We observed interspecific facilitation, where maize improves iron nutrition in intercropped peanut, faba bean enhances nitrogen and phosphorus uptake by intercropped maize, and chickpea facilitates P uptake by associated wheat from phytate-P. Furthermore, intercropping reduced the nitrate content in the soil profile as intercropping uses soil nutrients more efficiently than sole cropping.     

The role of maize root size in phosphorus uptake and productivity of maize/faba bean and maize/wheat intercropping systems

Interspecific root/rhizosphere interactions affect phosphorus (P) uptake and the productivity of maize/faba bean and maize/wheat intercropping systems. The aim of these experiments was to determine whether manipulation of maize root growth could improve the productivity of the two intercropping systems. Two near isogenic maize hybrids (the larger-rooted T149 and smaller-rooted T222) were intercropped with faba bean and wheat, under conditions of high- and low-P availability. The larger-rooted T149 showed greater competitive ability than the smaller-rooted T222 in both maize/faba bean and maize/wheat intercropping systems. The higher competitive ability of T149 improved the productivity of the maize/faba bean intercropping system in P-sufficient conditions. In maize/wheat intercropping systems, root growth, shoot biomass, and P uptake of maize were inhibited by wheat, regardless of the P-supply. Compared with T222, the larger-rooted T149 suffered less in the intercropping systems. The total biomass of the maize/wheat intercropping system was higher for wheat/T149 than for wheat/T222 under low-P conditions. These data suggested that genetic improvement of maize root size could enhance maize growth and its ability to compete for P resources in maize/faba bean and maize/wheat intercropping systems. In addition, depending on the P availability, larger maize roots could increase the productivity of intercropping systems.     

Effects of rhizobial inoculation, cropping systems and growth stages on endophytic bacterial community of soybean roots

Interspecific interactions and soil nitrogen supply levels affect intercropping productivity. We hypothesized that interspecific competition can be alleviated by increasing N application rate and yield advantage can be obtained in competitive systems. A field experiment was conducted in Wuwei, Gansu province in 2007 and 2008 to study intercropping of faba bean/maize, wheat/maize, barley/maize and the corresponding monocultures of faba bean (Vicia faba L.), wheat (Triticum aestivum L.), barley (Hordeum vulgare L.) and maize (Zea mays L.) with N application rates of 0, 75, 150, 225 and 300 kg N ha^?1. Total land equivalent ratios (TLER) were 1.22 for faba bean/maize, 1.16 for wheat/maize, and 1.13 for barley/maize intercropping over the 2-year study period. Maize was overyielding when intercropped with faba bean, but underyielding when intercropped with wheat or barley according to partial land equivalent ratios (PLER) based on grain yields of individual crops in intercropping and sole cropping. There was an interspecific facilitation between intercropped faba bean and maize, and interspecific competition between maize and either wheat or barley. The underyielding of maize was higher when intercropped with barley than with wheat. Fertilizer N alleviated competitive interactions in intercrops with adequate fertilizer N at 225 kg ha^?1. Yield advantage of intercropping can be acquired with adequate nitrogen supply, even in an intensive competitive system such as barley/maize intercropping. This is important when using intercropping to develop intensive farming systems with high inputs and high outputs.     

An overview of rhizosphere processes related with plant nutrition in major cropping systems in China

Rhizosphere processes of individual plants have been widely investigated since 1904 when the term “rhizosphere” was first put forward. However, little attention has been paid to rhizosphere effects at an agro-ecosystem level. This paper presents recent research on the rhizosphere processes in relation to plant nutrition in main cropping systems in China. In the peanut (Arachis hypogaea L.)/maize (Zea mays L.) intercropping system, maize was found to improve the Fe nutrition of peanut through influencing its rhizosphere processes, suggesting an important role of phytosiderophores released from Fe-deficient maize. Intercropping between maize and faba bean (Vicia faba L.) was found to improve nitrogen and phosphorus uptake in the two crops compared with corresponding sole crop. There was a higher land equivalent ratio (LER) in the intercropping system of maize and faba bean than the treatment of no root interactions between the two crops. The increased yield of maize intercropped with faba bean resulted from an interspecific facilitation in nutrient uptake, depending on interspecific root interactions of the two crops. In the rotation system of rice (Oryza sativa L.)-wheat (Triticum aestivum L.) crops, Mn deficiency in wheat was caused by excessive Mn uptake by rice and Mn leaching from topsoil to subsoil due to periodic cycles of flooding and drying. However, wheat genotypes tolerant to Mn deficiency tended to distribute more roots to deeper soil layer and thus expand their rhizosphere zones in the Mn-deficient soils and utilize Mn from the subsoil. Deep ploughing also helped root penetration into subsoil and was propitious to correcting Mn deficiency in wheat rotated with rice. In comparison, oilseed rape (Brassica napus L.) took up more Mn than wheat through mobilizing sparingly soluble soil Mn due to acidification and reduction processes in the rhizosphere. Thus, oilseed rape was tolerant to the Mn-deficient conditions in the rice-oilseed rape rotation. Oxidation reactions on root surface of rice also resulted in the formation of Fe plaque in the rice rhizosphere. Large amounts of Zn were accumulated on the Fe plaque. Zinc uptake by rice plants increased as Fe plaque formed, but decreased at high amounts of Fe plaque. It is suggested that to fine-tune cropping patterns and optimize nutrient management based on a better understanding of rhizosphere processes at an agro-ecosystem level is crucial for increasing nutrient use efficiency and developing sustainable agriculture in China.