地上和地下植食者互作对水稻氮分配及土壤活性氮的影响

植物茎叶和根系植食者虽然在空间上隔离,但二者的相互作用被认为是联系地上和地下部生态系统的基础。土壤氮素有效性通过植物影响植食者已得到大量研究的证实,但有关地上和地下部植食者互作对土壤氮素影响的研究却鲜有报道。以水稻褐飞虱和潜根线虫分别作为地上和地下的植食者,采用两因素交互试验设计,两个褐飞虱水平(未接种褐飞虱、接种褐飞虱),两个潜根线虫水平(未接种潜根线虫、接种潜根线虫),探讨二者的交互作用对水稻氮吸收和土壤活性氮(微生物生物量氮、可溶性有机氮以及铵态氮和硝态氮)的影响。结果表明,褐飞虱和潜根线虫相互抑制,二者的共存加剧了对水稻茎叶和根系生长的危害。褐飞虱未影响水稻茎叶和根系含氮量,而潜根线虫显著降低了水稻根系含氮量(p0.05)。褐飞虱和潜根线虫对土壤活性氮的影响表现出强烈的交互作用,在未接潜根线虫的处理中,褐飞虱显著提高了微生物生物量氮含量(p0.05),显著降低了硝态氮含量(p0.05);潜根线虫显著影响了微生物生物量氮含量和土壤活性氮总量。总之,褐飞虱和潜根线虫的相互抑制关系对土壤活性氮的影响格局较为复杂,相比褐飞虱,潜根线虫趋向于提高土壤活性氮水平,这可能影响与氮转化有关的土壤生态功能。     

Identification and Function Analysis of enolase Gene NlEno1 from Nilaparvata lugens (St?l) (Hemiptera:Delphacidae)

The enolase [EC 4.2.1.11] is an essential enzyme in the glycolytic pathway catalyzing the conversion of 2-phosphoglycerate (2-PGE) to phosphoenolpyruvate (PEP). In this study, a full-length cDNA encoding α-enolase was cloned from rice brown planthopper (Nilaparvata lugens) and is provisionally designated as NlEno1. The cDNA sequence of NlEno1 was 1,851 bp with an open reading frame (ORF) of 1,305 bp and encoding 434 amino acids. The deduced protein shares high identity of 80–87% with ENO1-like protein from Hemiptera, Diptera, and Lepidoptera speices. The NlEno1 showed the highest mRNA expression level in hemolymph, followed by fat body, salivary gland, ovaries and egg, and showed trace mRNA levels in testis. The mRNA of NlEno1 showed up-regulated level in virulent N. lugens population Mudgo, IR56 and IR42 when compared with TN1 population. Injection of double-stranded RNA (dsRNA) of NlEno1 into the adults significantly down-regulated the NlEno1 mRNA level along with decreased eggs and offspring. Moreover, injection of NlEno1-dsRNA decreased mRNA level of Vitellogenin (Vg) gene. These results showed that the NlEno1, as a key glycolytic enzyme, may play roles in regulation of fecundity and adaptation of N. lugens to resistant rice varieties.     

制霉菌素对褐飞虱取食行为、营养及共生菌数量的影响

为明确制霉菌素对褐飞虱取食行为、营养和共生菌数量的影响,用150、300和600 U/m L制霉菌素处理的水稻饲养褐飞虱,利用刺探电位图、高效液相色谱和镜检法监测其取食行为及体内必需氨基酸含量和类酵母共生菌(yeast-like symbiotes,YLS)数量。结果表明,取食300、600 U/m L制霉菌素处理水稻的褐飞虱3龄若虫体内YLS数量显著低于未处理水稻(对照),而150 U/m L处理与对照无显著差异;5龄若虫体内YLS数量随制霉菌素浓度的增加而显著降低,但600 U/m L与300 U/m L处理间差异不显著。褐飞虱于韧皮部吸食波出现次数在对照中显著多于制霉菌素处理,不同浓度间无明显差异;制霉菌素对褐飞虱的吸食时间影响不明显,但对其体内必需氨基酸含量影响较大,600 U/m L处理下均显著低于对照和其余处理,300 U/m L处理下除丙氨酸、脯氨酸、蛋氨酸和酪氨酸外均显著低于对照,150 U/m L处理下也均显著低于对照。表明制霉菌素能显著降低褐飞虱体内YLS数量和各种必需氨基酸含量,但对其吸食时间的影响不明显。     

蚜虫病原细菌的分离鉴定及其对褐飞虱的杀虫活性初探

为寻找有效防治褐飞虱的病原微生物,采用平板划线法和稀释涂布法,从感病死亡的蔷薇蚜虫体中分离筛选出一株对褐飞虱具有较高致病性的菌株JSA1;通过形态学特征、生理生化特性及16S r DNA序列分析,对菌株JSA1进行了初步鉴定;采用盆栽法和稻苗浸渍法,对其杀虫活性进行了测定。鉴定结果显示菌株JSA1为铜绿假单胞菌Pseudomonas aeruginosa。杀虫活性测定结果表明:菌株JSA1对褐飞虱具有杀虫作用,且致死率随菌液浓度的升高和作用时间的延长而提高,但致死率低于40%;菌株JSA1发酵液的上清液和沉淀破碎液对褐飞虱3龄若虫均具有致病力,72 h后的校正死亡率分别为30.72%和82.53%,表明铜绿假单胞菌JSA1在褐飞虱的生物防治方面具有一定的应用前景。     

褐飞虱IR56种群的致害特征

含Bph3抗褐飞虱基因的水稻品种自20世纪80年代在东南亚推广后,迄今仍对褐飞虱有较好的抗性,受到我国抗褐飞虱育种专家的重视。在IR56水稻(含Bph3基因)上连续40多代胁迫饲养获得褐飞虱IR56寄主种群,从褐飞虱的生长、发育、繁殖以及水稻品种的抗感反应(SSST法测定)两方面对该种群的致害性进行了研究。结果表明:1)在抗性水稻IR56上,褐飞虱IR56种群的羽化率、初羽化成虫体质量、成虫寿命、产卵量、蜜露排泄量、体质量增量等均较褐飞虱TN1种群显著增加,若虫历期则显著缩短;与感虫水稻TN1上的IR56种群或TN1种群相比,除成虫寿命、蜜露排泄量和体质量增量显著下降外,羽化率、产卵量、卵孵化率无显著差异,初羽化成虫体质量、若虫历期尽管差异显著但数值上较接近(其差值不及IR56水稻上IR56种群与TN1种群差值的1/2)。2)水稻品种苗期抗性反应显示IR56水稻对褐飞虱IR56种群的抗性级别为7级,明显弱于对褐飞虱TN1种群的3级,但略强于TN1水稻对褐飞虱IR56种群或TN1种群的9级。显然,与褐飞虱TN1种群相比,褐飞虱IR56种群对抗性水稻IR56有较强的致害能力,与IR56种群或TN1种群对TN1水稻的强致害水平尽管有一定的差距,但多数指标较为接近。含Bph3基因的水稻品种RathuHeenati对IR56种群抗性为1级,推测可能与该水稻品种存在Bph3以外的其他抗虫基因有关。     

类酵母共生菌中两个组氨酸合成基因在褐飞虱发育中的作用

褐飞虱[Nilaparvata lugens(Stl)]是我国水稻上的主要害虫,专一性吸食必需氨基酸缺乏的水稻筛管液。利用基因组和转录组数据,人们构建了褐飞虱及其体内类酵母共生菌(YLS)参与组氨酸等必需氨基酸的合成途径。在此基础上,本研究旨在通过基因克隆和RNA干扰研究YLS的His2和His6(分别命名为EdeHis2和EdeHis6)在褐飞虱生长、发育和存活中的作用。同源搜索和系统发育分析表明EdeHis2和EdeHis6均源于YLS基因组,与绿僵菌His2和His6高度同源并在系统发育树中形成一个簇,但在褐飞虱基因组中无同源基因。基因的时空表达分析表明,His2和His6在褐飞虱的各个龄期均表达且呈现一定的波动性,在褐飞虱脂肪体中的表达量均显著高于头、足、体壁和中肠。此外,在褐飞虱头和翅的基因组中未能扩增到目的片段,而在腹部能扩增出目的片段。分别注射外源双链RNA(dsEdeHis2或dsEdeHis6)后的第2、4、6天,EdeHis2的表达量分别显著下调45%~60%,EdeHis6下调27%~55%。dsEdeHis2和dsEdeHis6分别使褐飞虱的死亡率提高了8.3%和9.2%,雌、雄虫的若虫期历期延长了0.43d、0.33d和0.65d、0.36d,但均未达到显著水平。另外,注射dsEdeHis6的褐飞虱雌雄成虫翅畸形率分别为11%和13%,显著高于对照组。综上所述,源于YLS的EdeHis2和EdeHis6参与褐飞虱组氨酸的合成,与褐飞虱的存活、发育和翅发育相关。     

Antixenosis and Tolerance of Rice Genotypes Against Brown Planthopper

Nine genotypes were evaluated under greenhouse conditions for antixenosis and tolerance against brown planthopper(BPH, Nilaparvata lugens St?l). In antixenosis studies, proportion of insects settled on a test genotype in relation to the susceptible control TN1 was recorded, with significantly lower proportion of nymphs(55.22%–59.18%), adult males(60.33%–60.75%), and adult females(80.56%–79.26%) settled on RP2068-18-3-5 and Ptb33 in relation to those on TN1. Based on number of feeding sites, the test genotypes were ranked in order from the highest to the lowest as RP2068-18-3-5, Ptb33, MR1523, Rathu Heenati, Sinnasivappu, ARC10550, MO1, INRC3021 and TN1. The order was exactly reverse in terms of fecundity expressed as number of eggs laid per female. In tolerance studies, days to wilt, functional plant loss index and plant dry weight loss to BPH dry weight produced were recorded. RP2068-18-3-5, Rathu Heenati and Ptb33 performed better than the other test genotypes. These results helped in relative quantification of BPH resistance levels in the genotypes. RP2068-18-3-5, a new effective source of BPH resistance, can be used in resistance breeding after tagging of resistant genes/QTLs linked to different parameters of antixenosis and tolerance with selectable molecular markers.     

2个新培育水稻材料对褐飞虱的抗性机制研究

旨在筛选抗褐飞虱水稻材料,研究其对褐飞虱抗性机制。通过苗期抗性鉴定、成株期抗性鉴定筛选出了2份新培育的水稻材料,并对这2份水稻材料进行了忌避性、抗生性和耐害性的抗性试验。在苗期1105113表现为抗虫级别,1105096为高抗,在成株期1105096和1105113对褐飞虱均免疫;褐飞虱若虫和成虫均对1105096和1105113具有较强的忌避性;1105096和1105113对褐飞虱若虫存活率显著低于对照;1105096和1105113对褐飞虱的耐害性较弱,抗生性较强。这2个水稻材料在苗期和成株期均对褐飞虱具有良好的抗性效果。同时,这2个水稻材料对褐飞虱兼具有忌避性、抗生性和耐害性,其中抗生性作用较强,耐害性作用较弱,均可用于培育抗褐飞虱水稻品种。     

烯啶虫胺对水稻褐飞虱田间控制效果

田间试验结果表明:烯啶虫胺对水稻褐飞虱具有较好的速效性、持效性,药后3 d,50%烯啶虫胺WSG 22.5~52.5g/hm2对褐飞虱的防治效果为84.70%~90.81%;药后14~21 d,50%烯啶虫胺WSG 37.5~52.5 g/hm2对褐飞虱仍有89.84%~96.06%的防治效果。因此,从防治效果和防治成本的角度考虑,50%烯啶虫胺WSG防治水稻褐飞虱的适宜用量为37.5~52.5 g/hm2,防治适期在褐飞虱若虫高峰期,用水量不低于750 kg/hm2。