花生抗黄曲霉菌侵染和产毒机制研究进展

黄曲霉菌（Aspergillus flavus L.）侵染花生导致黄曲霉毒素（Aflatoxin）污染是影响花生食用安全性和限制花生产业发展的重要因素,其污染抗性分为抗侵染和抗产毒两种类型。抗性机制主要包括形态机制、生理机制和分子机制。种皮中决定花生抗侵染的主要因素为蜡质层厚度、栅栏细胞排列密集程度及有无裂纹等特征。种子中单宁酸、胰蛋白酶抑制剂、白藜芦醇等与花生抵抗黄曲霉菌侵染及毒素合成有关。病程相关蛋白基因、转录因子基因、脂氧合酶基因等均参与了花生抵抗黄曲霉菌侵染的过程。随着生物技术的迅猛发展,将来可利用全基因关联分析和多组学结合的方法,从基因调控网络水平上开展花生黄曲霉抗性研究,在更深层次上阐明花生黄曲霉抗性机制。     

收获前花生黄曲霉综合预防控制技术

根据花生收获前黄曲霉感染因素,探讨了收获前栽培管理、病虫害防治和遗传控制等一系列措施,综合预防控制花生黄曲霉毒素污染.     

花生抗黄曲霉大果种质的创制与鉴定

培育和种植抗黄曲霉品种是防控花生黄曲霉毒素污染最为经济有效的途径,而黄曲霉抗性与高产的矛盾一直是花生抗黄曲霉育种的障碍.本研究以抗黄曲霉花生种质J11与高产品种中花16为亲本构建的重组自交系群体(Recombined inbreed lines,RILs)为材料,进行黄曲霉侵染和产毒抗性鉴定,探讨抗性与高产(大果)性状...     

黄曲霉对花生和玉米的侵染研究

黄曲霉(Aspergillus flavus)极易侵染花生、玉米等多种农作物,并且在侵染后产生具有致癌、致畸、致突变作用的有毒代谢产物——黄曲霉毒素.黄曲霉的侵染和黄曲霉毒素污染不仅发生在作物生长环节,而且在收获、干燥、储运等过程也会发生,严重威胁农产品消费安全和人畜生命健康.因此,解析黄曲霉对粮油作物的侵染过程及侵染...     

山东省花生产区土壤和荚果中黄曲霉菌及其毒素污染状况调查

为了解2018年山东省花生主产区土壤及收获后花生荚果黄曲霉菌及黄曲霉毒素的污染情况,在烟台、青岛、临沂、泰安、枣庄及菏泽等花生主要产区进行土壤、花生荚果的采样,对土壤、荚果果壳、花生籽仁的黄曲霉菌检出率进行统计并对籽仁中黄曲霉毒素的含量进行测定。结果表明：不同产区土壤中黄曲霉菌的检出率为3.33%~33.33%;花生果壳中黄曲霉菌的检出率为10.89%~27.78%;花生籽仁中黄曲霉菌的检出率为3.11%~11.56%;花生籽仁中黄曲霉毒素的浓度为5.01~26.80 μg/kg。数据分析得出：花生籽仁中黄曲霉毒素的含量与土壤中黄曲霉菌的检出率呈极强的相关性;与花生果壳和籽仁中黄曲霉菌的检出率呈中等程度相关。本研究对解析花生种植区黄曲霉菌及黄曲霉毒素污染发生原因,精准预警与防控,提高农产品质量安全有重要意义。     

广东花生产区土壤黄曲霉产毒菌落分布初步研究

采用稀释倒平板法对广东10个花生主产地土壤中黄曲霉的数量以及分离菌株的产毒能力进行了分析。结果表明：全省各地花生种植土壤中黄曲霉的数量存在较大的差异,其中清远市土壤中的黄曲霉数量最多,达160 cfu/g干土壤,惠州和广州地区次之,均为100 cfu/g干土壤,汕头地区最少,在该地区抽取的土壤样品中未分离出黄曲霉菌株。在分离出来的黄曲霉菌株中,广州地区有10株产毒菌株、清远地区17株、惠州地区16株、云浮地区8株、河源地区8株。     

湖北省典型花生种植区土壤中黄曲霉菌分布及产毒力研究

为掌握湖北省花生种植区土壤中黄曲霉菌的分布和产毒特征,从罗田、红安、钟祥、襄阳四个典型花生种植区采集土壤样品40份,并进行黄曲霉菌分离、鉴定和产毒力研究。研究结果表明:湖北省不同花生种植区共分离鉴定到黄曲霉菌51株,土壤中黄曲霉菌落数为127.5cfu/g。不同种植区土壤中黄曲霉菌分布存在显著差异,钟祥土壤中黄曲霉菌落数最高,罗田最低;鉴定获得黄曲霉菌株中产毒菌株占96%,产毒量范围0～227.81μg/L,不产毒菌株占4%;产毒菌株分为只产AFB_1、产AFB_1+AFB_2、产AFB_1+AFB_2+AFG_1和产AFB_1+AFB_2+AFG_1+AFG_2毒素4种类型,其中以产AFB_1+AFB_2的菌株占比最高,为65%;不同种植区黄曲霉菌株产毒力研究发现,钟祥每克土壤中黄曲霉菌产AFB_1的量最高,达11 679.70μg/L。本研究可为湖北花生黄曲霉毒素污染预警和防控提供理论依据。     

具有化感作用的植物能增强花生对黄曲霉毒素的抗性（英文）

研究三裂叶蟛蜞菊、香根草、香茅对黄曲霉的化感作用。结果表明,三种植物的水提液均能显著抑制黄曲霉的生长。在培养黄曲霉的培养基中加入上述三种植物的水提液后,培养基上黄曲霉的菌落数显著减少,平均菌落直径变小。利用这三种植物的水提液浸泡被黄曲霉感染的花生种子,其发芽率显著高于清水对照处理;显示这三种植物的水提液能够缓解黄曲霉素对花生种子发芽的抑制作用。在没有黄曲霉侵染的条件下,虽然这三种植物的水提液会降低正在发芽的花生种子某些水解酶和保护酶的活性;但在黄曲霉侵染后,这三种植物水提液浸泡过的花生种子水解酶和保护酶的活性却显著高于对照处理,导致处理种子具有更高的呼吸速率、种子活力和发芽率。在对照土壤里,受黄曲霉侵染的种子长出的花生植株长势很差。分别将三种化感植物剪碎、加入土壤中混匀,平衡一段时间;在这些土壤上种植受黄曲霉侵染的种子长出的花生发现能够有效降低黄曲霉素对花生生长的抑制作用。与对照相比,处理土壤上生长的受黄曲霉侵染的花生植株具有更高的根系活力、叶绿素含量、净光合速率、硝酸还原酶和谷氨酰胺合成酶活性,因而具有更高的株高、叶片氮素含量和干物量。     

花生抗青枯病种质对黄曲霉菌产毒的抗性反应

通过人工接种和毒素测定,对抗、感青枯病的花生种质进行了黄曲霉菌产毒抗性评价.结果表明:1)抗青枯病花生种质之间存在着黄曲霉产毒抗性的显著差异,品种间毒素含量高低相差近10倍,鉴定出抗黄曲霉产毒种质6个;2)2种抗病性无相关性;3)获得具有直接生产利用价值的兼抗青枯病和抗黄曲霉产毒的花生材料1个.     

中国西南花生产区黄曲霉菌分布、产毒力及花生黄曲霉毒素污染

为探明中国西南花生产区黄曲霉菌分布、产毒力及产后花生黄曲霉毒素污染情况,从云南广南、西藏察隅、四川蓬安采集177份花生、土壤样品,共分离鉴定出黄曲霉菌206株,分析其分布及产毒特征,并调查71份产后花生样品黄曲霉毒素污染情况。结果表明,四川蓬安花生产地黄曲霉菌分布最广,数量最多,其检出率和菌落数分别为50.0%、212.0 CFU/g,西藏察隅黄曲霉菌分布最少（18.1%,52.8 CFU/g）。菌株产毒力研究结果表明,88.6%的菌 株能产生黄曲霉毒素,产毒类型以B族毒素为主,尤其是AFB1,其含量在0～8500μg/L,平均产毒能力排序为：云南广南（4393.4μg/L）>西藏察隅（1991.0μg/L）>四川蓬（1259.3μg/L）;不产毒菌株占11.4%,均来自西藏察隅。对产后花生黄曲霉毒素污染研究发现,西南地区花生黄曲霉毒素污染较轻,阳性样品检出率为7.0%,AFB1含量在0~7.2 μg/kg;四川蓬安花生样品AFB1检出率为4.2%,含量0~7.2 μg/kg,云南广南AFB1检出率为16.7%,含量0~2.1 μg/kg, 西藏察隅样品未检出AFB1。西南产区黄曲霉菌检出率越高、菌落数越多,产后花生黄曲霉毒素污染越严重。     

Field evaluation of resistance to aflatoxin accumulation in maize inbred lines in Kenya and South Africa

Aflatoxin, a carcinogenic toxin, is produced mainly by Aspergillus flavus and Aspergillus parasiticus. Contamination of maize (Zea mays L.) grain by these fungi occurs before harvest, and the easiest strategy to prevent this is to develop/use maize varieties resistant to Aspergillus spp. and aflatoxin accumulation. The objective of this investigation was to identify potential sources of resistance among 23 maize inbred lines (13 obtained from the MAIZE Competitive Grants Initiative, International Maize and Wheat Improvement Centre and 10 from Agricultural Research Council, South Africa). The inbred lines were planted in a randomized complete-block design at two locations each in Kenya and South Africa. Maize ears were inoculated at silking with three toxigenic strains of A. flavus. The inoculated ears in each plot were harvested at 12–18% moisture, dried, and visually assessed for Aspergillus ear rot (AER). Aflatoxin concentration in the kernels was determined using liquid chromatography–tandem mass spectrometry. Significant variation for both AER and aflatoxin concentration existed among the inbred lines at both locations in Kenya and one location in South Africa. Combined analysis revealed a significant (p < 0.001) lines × locations interaction for both AER and aflatoxin concentration. Higher incidences of AER (0–86.0%) and aflatoxin concentration (0.21–6.51 µg/kg) were recorded at Kiboko in Kenya than at the other three locations. A stronger genetic correlation (r_G = 0.936, p < 0.0001) between the AER and aflatoxin concentration was recorded in Potchefstroom than at the other three locations. Repeatability of aflatoxin concentration was high at Kiboko (0.87) and Potchefstroom in South Africa (0.74). Three inbred lines, CML247, CML444, and CML495, emerged as potentially useful sources of resistance to AER and aflatoxin accumulation as they showed low levels of aflatoxin contamination in both localities in Kenya and in South Africa.     

Association between aflatoxin contamination and drought tolerance traits in peanut

The current study investigates the association between drought tolerance traits and aflatoxin contamination in peanut grown under long-term drought. Two field experiments were conducted at Khon Kaen University, Thailand using a split–split plot design with three drought stress levels as main plots, 11 genotypes as sub-plots, and two soil inoculations of Aspergillus flavus treatments as sub-sub-plots. The effects of temperature, soil moisture and A. flavus population on kernel colonization and aflatoxin contamination, and drought tolerance traits viz. specific leaf area (SLA) and root length density (RLD) were measured. The results demonstrated that elevated soil temperatures and reduced soil moisture, favored aflatoxin production. Drought in combination with higher levels of A. flavus inoculum load in the soil resulted in an increase in the fungal populations in the soil which in turn resulted in increased kernel colonization and subsequent aflatoxin contamination. A combination of SLA and RLD, and kernel colonization had a significant influence on aflatoxin contamination under drought conditions in both seasons (r = 0.73** and 0.76**). The results revealed that drought tolerance traits (SLA and RLD) could be contributing to resistance to aflatoxin contamination suggesting that a combination of SLA, RLD and kernel colonization could be used as selection criteria in selecting parents for aflatoxin resistance.     

花生品质性状与抗黄曲霉侵染关系的研究

以对黄曲霉具有不同抗性的花生品种为材料,通过人工黄曲霉接种测定和品质性状的分析,探讨花生品质性状与抗黄曲霉的关系。结果表明,花生种子胰蛋白酶抑制剂活性的高低(PTI值)与品种黄曲霉侵染率呈显著负相关;而种子可溶性糖、蔗糖和蛋白质含量与品种抗黄曲霉侵染能力无关,花生种子胰蛋白酶抑制剂活性的高低可以作为花生抗黄曲霉育种选育的品质性状标记。     

花生编码磷脂酶D的类PLD基因的鉴定和部分测序

花生工业界认为收获前花生黄曲霉素养素的污染是全世界花生工业界面临的最严峻的挑战，干旱胁迫是加重花生花曲霉真菌侵染和毒素污染最重要的环境因素。选育花生抗性品种将使美国花生工业处于优势地位。在这一研究报告中，我们鉴定出了一个新的类PLD基因，它编码磷脂酶D。在植物体中，这个酶是负责干旱诱导降解细胞膜磷脂的主要酶，克隆的PLD1片段有1069个核甘酸对长。推导的氨基酸序列与已知的PLD基因有很高的同一性，包括相似的保守序列特征，比如两个HXKXXXXD基元，对花生PLD基因特性需要从遗传和生理上作进一步研究，以便更好地理解这个基因的功能及其与花生耐干旱性的关系。     

花生荚壳抗黄曲霉菌侵染的鉴定方法研究及抗性种质发掘

        黄曲霉毒素污染是影响花生食用安全性和制约产业发展的重要因素。荚壳是花生抵御黄曲霉菌侵染的第一道防线,为建立花生荚壳抗黄曲霉侵染的鉴定方法,本研究利用强侵染黄曲霉菌AF2202接种花生荚果,通过对不同接种菌浓度和培养时间的比较分析,发现接种浓度为2 ×106孢子/mL、培养7d的组合方案可以有效区分花生荚壳对黄曲霉菌侵染的抗性。利用所建立的鉴定方法对276份遗传变异丰富的花生核心种质材料进行接种鉴定,进一步证明了这一方法鉴定花生荚壳抗性的有效性和实用性,初步发掘出2份具有荚壳抗性的特异花生种质。     

Diallel analysis of aflatoxin accumulation in maize

Since its discovery in numerous feedstuffs, aflatoxin, a carcinogenic compound produced by the fungus Aspergillus flavus Link ex Fries, has caused much concern among consumers and producers alike. This toxin poses a serious economic threat to maize (Zea mays L.) producers of the southeastern and midwestern regions of the United States. Efforts to identify maize germplasm that is resistant to aflatoxin accumulation and to investigate the genetic basis of this resistance have been undertaken at numerous research institutions. The objectives of this study were to (1) evaluate aflatoxin accumulation in grain harvested from maize inbred lines and a diallel cross among these lines, (2) determine the importance of general and specific combining abilities in inheritance of resistance to aflatoxin accumulation, and (3) estimate general and specific combining ability effects associated with resistance to aflatoxin accumulation in the inbred lines and crosses among them. Eight inbred lines and a diallel cross of the maize lines were inoculated with an A. flavus spore suspension 12–14 d after silk emergence. Following harvest, aflatoxin content was determined from samples of grain. Statistical analyses performed using SAS general linear models (GLM) and DIALLEL-SAS indicated that general and specific combining ability were significant sources of variation in the inheritance of resistance to aflatoxin accumulation. The inbred line Mp313E, which was developed and released as a source of resistance to aflatoxin contamination, had significantly lower aflatoxin accumulation than other lines. Mo18W exhibited excellent general combining ability for reduced aflatoxin accumulation when crossed with the other lines. Both Mp313E and Mo18W could be useful in breeding programs to develop aflatoxin-resistant maize hybrids. Mp339, SC212M, and Ab24E demonstrated aflatoxin susceptibility as both inbreds and in single crosses.     

花生种质资源对黄曲霉菌侵染和产毒的抗性鉴定

对不同种子成分的花生种质资源对黄曲霉菌侵染和产毒的抗性进行了鉴定,结果表明,高含油量、高蛋白质含量和高油酸含量资源对黄曲霉菌侵染和产毒的抗性较差.通过对抗黄曲霉侵染和产毒能力不同的花生种质资源的种子成分的分析表明,高抗黄曲霉菌侵染和产毒资源的亚油酸含量较高.相关分析表明,不同花生品种对黄曲霉菌的侵染抗性与种子大小和油酸含量呈显著负相关,与出仁率和亚油酸含量呈显著正相关;不同花生品种对黄曲霉菌产毒的抗性与含油量呈显著负相关.通过鉴定和筛选,发掘出2份优质抗病资源.     

Effect of non-aflatoxigenic strains of Aspergillus flavus on aflatoxin contamination of pre-harvest peanuts in fields in China

Aflatoxin contamination of peanuts is one of the most concerns in peanut production in China. Applying non-aflatoxigenic Aspergillus flavus strains, based on competitive exclusion, has been proved to be a promising strategy to reduce aflatoxin contamination in pre-harvest peanuts. Two non-aflatoxigenic A. flavus strains collected in China, which have been proved effectively reducing aflatoxin in the laboratory, were mixed with high aflatoxin producer to the soil in peanut growing season. The two non-aflatoxigenic strains significantly (P ​< ​0.05) reduced aflatoxin contamination in peanut kernels under both normal and drought stresses in two fields. Compared to control, the total aflatoxin (sum of aflatoxin B₁ and B₂) was reduced 26.7–99.12% in field 1, and 84.96–99.33% in field 2. The aflatoxin was reduced 84.96–99.33% under drought stress in two fields. The present study indicated the non-aflatoxigenic A. flavus strains could be potential biocontrol agents for reducing aflatoxin contamination under field condition.     

Fungal species isolated from peanuts in major Kenyan markets: Emphasis on Aspergillus section Flavi

A survey was conducted in Nairobi, Nyanza and Western provinces in Kenya between March and July 2009 with 1263 peanut products sampled out of which 705 samples underwent microbial analysis. The study aimed at determining the incidence of fungal species – emphasis on Aspergillus section Flavi – associated with peanut products. A 0.5 kg representative sample was obtained from each surveyed vendor and the colony forming units (CFU) of fungal species determined. The samples were also analyzed for total aflatoxin level while isolates of Aspergillus flavus and Aspergillus parasiticus were screened for production of aflatoxin B1, B2, G1 and G2. Eight fungal species were detected in the samples and were in decreasing order of CFU/g of sample: A. flavus S-strain (467), A. flavus L-strain (341), Penicillium spp. (326), Aspergillus niger (156), Aspergillus tamari (27), Aspergillus alliaceus (21), A. parasiticus (10), and Aspergillus caelatus (5). The overall incidence of A. flavus S-strain in samples from Nairobi was 92 and 1425% higher than samples from Nyanza and Western regions, respectively. The combined incidence of A. flavus and A. parasiticus was varied significantly (p ≤ 0.05) with peanut product: peanut flour (69%), shelled raw peanuts (53%), spoilt peanuts (49%), boiled podded peanuts (45%), podded peanuts (39%), peanut butter (31%), fried peanuts (22%) and roasted peanuts (20%). Seventy three percent of A. flavus and A. parasiticus isolates produced at least one of the aflatoxin types, with 66% producing aflatoxin B1. The total aflatoxin level among peanut products ranged from 0 to 1629 μg/g; and there was a positive correlation (r = 0.2711) between the incidence of A. flavus and A. parasiticus, and total aflatoxin level. The high incidence of aflatoxin producing fungi in peanuts traded in Kenyan markets implies a risk of aflatoxin contamination, highlighting the need for stakeholders to promote sound practices at all stages of the peanut value chain in order to minimize market access by non-complying products.     

Diallel analysis of fumonisin accumulation in maize

Contamination of maize (Zea mays L.) grain with aflatoxin, produced by the fungus Aspergillus flavus Link: Fries, or fumonisin, produced by the fungus Fusarium verticillioides (Saccardo) Nirenberg (Syn F. moniliforme Sheldon), greatly reduces its value and marketability. Host resistance to fungal infection and mycotoxin accumulation is considered a highly desirable way to reduce losses of both aflatoxin and fumonisin. Maize germplasm lines with resistance to aflatoxin accumulation have been developed in Mississippi. Three of the aflatoxin-resistant lines and five additional lines were used as parents to produce a diallel cross. The diallel cross was evaluated for fumonisin accumulation in field tests conducted in Mississippi in 2007 and 2008. General combining ability (GCA) was a highly significant source of variation in both 2007 and 2008, but specific combining ability (SCA) was not significant. GCA effects for reduced fumonisin levels were highly significant for Mp715, Mp717, and GA209. Mp715 and Mp717 were developed and released as sources of resistance to aflatoxin contamination and exhibit resistance as inbred lines and in hybrid combinations. These lines should be useful in developing maize lines and hybrids with resistance to both fumonisin and aflatoxin accumulation in grain.