CN106353162A - 一种用于禾草内生真菌检测的无毒染色剂 - Google Patents
一种用于禾草内生真菌检测的无毒染色剂 Download PDFInfo
- Publication number
- CN106353162A CN106353162A CN201610674349.2A CN201610674349A CN106353162A CN 106353162 A CN106353162 A CN 106353162A CN 201610674349 A CN201610674349 A CN 201610674349A CN 106353162 A CN106353162 A CN 106353162A
- Authority
- CN
- China
- Prior art keywords
- detection
- endogenetic fungus
- stain
- dogstail
- nontoxic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000233866 Fungi Species 0.000 title claims abstract description 71
- 231100000252 nontoxic Toxicity 0.000 title claims abstract description 38
- 230000003000 nontoxic effect Effects 0.000 title claims abstract description 38
- 244000025254 Cannabis sativa Species 0.000 title claims abstract description 7
- 239000003086 colorant Substances 0.000 title abstract 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 22
- XOSXWYQMOYSSKB-LDKJGXKFSA-L water blue Chemical compound CC1=CC(/C(\C(C=C2)=CC=C2NC(C=C2)=CC=C2S([O-])(=O)=O)=C(\C=C2)/C=C/C\2=N\C(C=C2)=CC=C2S([O-])(=O)=O)=CC(S(O)(=O)=O)=C1N.[Na+].[Na+] XOSXWYQMOYSSKB-LDKJGXKFSA-L 0.000 claims abstract description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000004310 lactic acid Substances 0.000 claims abstract description 11
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 11
- 230000002538 fungal effect Effects 0.000 claims abstract description 8
- 239000002904 solvent Substances 0.000 claims abstract description 7
- 238000010438 heat treatment Methods 0.000 claims abstract 2
- 238000001514 detection method Methods 0.000 claims description 66
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 10
- 239000012153 distilled water Substances 0.000 claims description 6
- 238000009835 boiling Methods 0.000 claims description 4
- 239000002689 soil Substances 0.000 claims description 4
- 239000008399 tap water Substances 0.000 claims description 4
- 235000020679 tap water Nutrition 0.000 claims description 4
- 239000003513 alkali Substances 0.000 claims description 3
- 230000001580 bacterial effect Effects 0.000 claims description 3
- 235000016709 nutrition Nutrition 0.000 claims description 3
- 230000035764 nutrition Effects 0.000 claims description 3
- 238000007790 scraping Methods 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- 210000000481 breast Anatomy 0.000 claims 1
- 239000011521 glass Substances 0.000 claims 1
- 239000000203 mixture Substances 0.000 claims 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 abstract description 22
- 210000002421 cell wall Anatomy 0.000 abstract description 5
- 241000282414 Homo sapiens Species 0.000 abstract description 3
- 238000004040 coloring Methods 0.000 abstract 2
- 230000000694 effects Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 7
- 238000004043 dyeing Methods 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 238000000034 method Methods 0.000 description 5
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N Aniline Chemical compound NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 241000221752 Epichloe Species 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 208000009304 Acute Kidney Injury Diseases 0.000 description 2
- 206010019233 Headaches Diseases 0.000 description 2
- 206010037423 Pulmonary oedema Diseases 0.000 description 2
- 208000033626 Renal failure acute Diseases 0.000 description 2
- 201000011040 acute kidney failure Diseases 0.000 description 2
- 208000012998 acute renal failure Diseases 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 208000002173 dizziness Diseases 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- MURGITYSBWUQTI-UHFFFAOYSA-N fluorescin Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC=C(O)C=C2OC2=CC(O)=CC=C21 MURGITYSBWUQTI-UHFFFAOYSA-N 0.000 description 2
- 231100000869 headache Toxicity 0.000 description 2
- 238000003119 immunoblot Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 231100000614 poison Toxicity 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 208000005333 pulmonary edema Diseases 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 206010001580 Albuminuria Diseases 0.000 description 1
- 241000235349 Ascomycota Species 0.000 description 1
- 241000124150 Balansia Species 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 206010017533 Fungal infection Diseases 0.000 description 1
- 244000283207 Indigofera tinctoria Species 0.000 description 1
- 208000031888 Mycoses Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000190478 Neotyphodium Species 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 241000209504 Poaceae Species 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 208000004756 Respiratory Insufficiency Diseases 0.000 description 1
- 206010047513 Vision blurred Diseases 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 231100000570 acute poisoning Toxicity 0.000 description 1
- 230000005875 antibody response Effects 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 231100000739 chronic poisoning Toxicity 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 239000011280 coal tar Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 231100000206 health hazard Toxicity 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 206010022694 intestinal perforation Diseases 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 231100000636 lethal dose Toxicity 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 201000004193 respiratory failure Diseases 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000037384 skin absorption Effects 0.000 description 1
- 231100000274 skin absorption Toxicity 0.000 description 1
- 239000011877 solvent mixture Substances 0.000 description 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明提供了一种用于禾草内生真菌检测的无毒染色剂包括以下成分:苯胺蓝、乳酸和溶剂,该无毒染色剂不含对人体和环境有危害的苯酚,同时还通过在染色过程中将玻片在酒精灯火焰上简单加热,提高了染色效果,便于区别内生真菌菌丝和细胞壁。
Description
技术领域
本发明涉及真菌检测技术领域,特别地,涉及一种用于禾草内生真菌检测的无毒染色剂。
背景技术
禾草内生真菌(grass endophyte)可以在禾草体内生存大部分时间或者全部时间都寄生在禾草体内,但却不会引起禾草外部显示任何病害症状(Siegel et al.,1987)。广泛包括真茵子囊菌门的香柱菌属(Epichloe),瘤座菌属(Balansia)VA菌根菌、真菌半知菌类的一些属(Clay,1991),其中研究较多的是与禾本科草类形成一种共生的香柱菌属及其无性态Neotyphodium内生真菌(黄龙益和黄小龙,2008)
内生真菌具有寄主特异性,分离自不同的寄主的内生真菌具有不同的特征,这与寄主被真菌侵染的过程有关(Toti et al.,1992)。检测禾草内生真菌的方法有很多,自上个世纪70年代起,国内外对内生真菌的检测方法就在不断完善,简化,精确。通过对内生真菌进行分离检测,从形态学上鉴定内生真菌的形态从而鉴定属种,但是相对来说比较费时,检测周期长,因环境等不可控因素造成的菌种保存程度等也会对菌种的鉴定产生一定的影响。利用组织染色(Bacon et al.,1977),在显微境下观察组织与组织培养物,其快速、直观,适用于少量样本的检测,但是当检测样本组织内的内生真菌含量少时,依靠染色镜检则很难检测出来。除此之外还有荧光蛋白染色法(GFP),扫描、透射电子显微法等其他显微镜检测方法(Christensen et al.,1997),相比光镜下检测染色组织过程更复杂也更细致,但是成本相对来说偏高。适用于大量样本的检测方法有免疫印迹法(immunoblot)(Gwinn etal,1991),酶联免疫吸附法(ELISA)(Musgrave,1984)等,通过利用免疫学上的特点,利用内生真菌特异性来鉴定不同的禾草内生真菌种类,但这要求检测的样品质量很高,如果控制不当,植物蛋白就会与抗体反应从而影响到结果。目前,显微观察法仍然是最常用的检测方法,该检测方法简叙如下:
一、染色剂配方(聂立影,2004;张盼盼,2010;李春杰,2008):
1号溶液:5%NaOH和1%苯胺蓝混合溶液
2号溶液:100mL乳酸,l00mL甘油,l00mL苯酚,800mg苯胺兰,500mL蒸馏水
二、内生真菌检测方法
种子中内生真菌的检测:取200粒种子放入50mL烧杯中,室温下用1号溶浸泡24h,用自来水冲洗干净后,去除种子的颖片,再转入装有2号溶液的试管中,在沸水浴中加热l0min,保证充分染色,取一粒种子置于一滴蒸馏水的载玻片上,压片,在40倍体视显微镜下镜检,观察细胞间的菌丝情况。
叶鞘中内生真菌的检测:将禾草分蘖剪下,用刀片和镊子剥取分蘖叶鞘基部的内表皮组织,置于载玻片上,滴一滴苯胺蓝溶液,待充分染色后,在40倍显微镜下进行镜检,观察细胞间的菌丝情况。
苯酚,又名石炭酸、羟基苯,是德国化学家龙格(Runge F)于1834年在煤焦油中发现的,是最简单的酚类有机物,一种弱酸。常温下为一种无色晶体,有毒。
1)健康危害:苯酚对皮肤、粘膜有强烈的腐蚀作用,可抑制中枢神经或损害肝、肾功能。急性中毒:吸入高浓度蒸气可致头痛、头晕、乏力、视物模糊、肺水肿等。误服引起消化道灼伤,出现烧灼痛,呼出气带酚味,呕吐物或大便可带血液,有胃肠穿孔的可能,可出现休克、肺水肿、肝或肾损害,出现急性肾功能衰竭,可死于呼吸衰竭。眼接触可致灼伤。可经灼伤皮肤吸收经一定潜伏期后引起急性肾功能衰竭。慢性中毒:可引起头痛、头晕、咳嗽、食欲减退、恶心、呕吐,严重者引起蛋白尿。可致皮炎。人口服致死量报道不一,LD为2~15g,或MLD为140mg/kg,14g/kg。国外报道酚液污染皮肤面积为25%,10分钟死亡,血酚为0.74mmlo/L。
2)环境危害:对环境有严重危害,对水体和大气均可造成污染。
3)燃爆危险:该品可燃,高毒,具强腐蚀性。
该检测方法中2号溶液中含有苯酚,不利于工作人员身体健康,同时废液易污染大气和土壤,故有必要研发一种新的用于禾草内生真菌检测的无毒染色剂。
发明内容
本发明目的在于提供一种用于禾草内生真菌检测的无毒染色剂,以解决技术问题。
一种用于禾草内生真菌检测的无毒染色剂,包括以下成分:苯胺蓝、乳酸和溶剂。
优选的,所述的无毒染色剂用于种子内生真菌检测时,溶剂为水。
优选的,所述的无毒染色剂用于种子内生真菌检测时,包括以下成分:0.325g苯胺蓝,100ml水,50ml 85%的乳酸。
优选的,所述的无毒染色剂用于叶鞘内生真菌检测或茎髓内生真菌检测时,溶剂为丙三醇和水的混合物。
优选的,所述的无毒染色剂用于叶鞘内生真菌检测或茎髓内生真菌检测时,包括以下成分:1份乳酸,2份丙三醇,1份蒸馏水,质量分数为0.05%的苯胺蓝。
所述的用于禾草内生真菌检测的无毒染色剂用于种子内生真菌检测法,包括以下步骤:
将100粒种子放入20ml烧杯中,倒入5%的氢氧化钠溶液浸泡过夜,自来水冲洗掉碱液,加入染色剂,加热至沸腾再冷却,取出一粒种子放在滴有苯胺蓝无毒染色剂的载玻片上,盖上盖玻片,轻轻按压种子,在物镜10倍和40倍下观察菌丝情况。
所述的用于禾草内生真菌检测的无毒染色剂用于叶鞘内生真菌检测法,包括以下步骤:将带有内生真菌的禾草种子播种于营养土中,取6周龄的幼苗进行带菌率检测,用镊子和解剖刀撕取叶鞘部分内表皮,放于载玻片上加染色剂进行染色,盖上盖玻片,并在酒精灯火焰上加热5-8秒,静置片刻后在显微镜下观察内生真菌菌丝。
所述的用于禾草内生真菌检测的无毒染色剂用于茎髓内生真菌检测法,包括以下步骤:用解剖刀将禾草成株茎秆纵向剖开,刀尖刮取茎杆髓质放于载玻片上,用染色剂进行染色,盖上盖玻片,并在酒精灯火焰上加热5-8秒,静置片刻后在显微镜下观察内生真菌菌丝。
本发明的染色剂中不需要添加苯酚,在染色过程中仅需要将玻片在酒精灯火焰上加热数秒即可;同时染色效果好,原技术容易将细胞壁染色,影响观察,而本发明的染色剂只能将内生真菌的菌丝染色,不会对细胞壁染色,便于区别内生真菌菌丝和细胞壁。
本发明具有以下有益效果:本发明的用于禾草内生真菌检测的无毒染色剂,包括以下成分:苯胺蓝、乳酸和溶剂,该无毒染色剂不含对人体和环境有危害的苯酚,同时还通过在染色过程中将玻片在酒精灯火焰上简单加热,提高了染色效果,便于区别内生真菌菌丝和细胞壁。
除了上面所描述的目的、特征和优点之外,本发明还有其它的目的、特征和优点。下面将对本发明作进一步详细的说明。
具体实施方式
以下对本发明的实施例进行详细说明,但是本发明可以根据权利要求限定和覆盖的多种不同方式实施。
实施例1
一种用于禾草内生真菌检测的无毒染色剂,包括以下成分:0.325g苯胺蓝,100ml水,50ml 85%的乳酸;
所述的用于禾草内生真菌检测的无毒染色剂用于种子内生真菌检测法,包括以下步骤:
将100粒种子放入20ml烧杯中,倒入5%的氢氧化钠溶液浸泡过夜,自来水冲洗掉碱液,加入染色剂,加热至沸腾再冷却室温,取出一粒种子放在滴有苯胺蓝无毒染色剂的载玻片上,盖上盖玻片,轻轻按压种子,在物镜10倍和40倍下观察菌丝情况。
实施例2
一种用于禾草内生真菌检测的无毒染色剂,包括以下成分:1份乳酸,2份丙三醇,1份蒸馏水,质量分数为0.05%的苯胺蓝。
所述的用于禾草内生真菌检测的无毒染色剂用于叶鞘内生真菌检测法,包括以下步骤:将带有内生真菌的禾草种子播种于营养土中,取6周龄的幼苗进行带菌率检测,用镊子和解剖刀撕取叶鞘部分内表皮,放于载玻片上加染色剂进行染色,盖上盖玻片,并在酒精灯火焰上加热7秒,静置片刻后在显微镜下观察内生真菌菌丝。
实施例3
一种用于禾草内生真菌检测的无毒染色剂,包括以下成分:1份乳酸,2份丙三醇,1份蒸馏水,质量分数为0.05%的苯胺蓝。
所述的用于禾草内生真菌检测的无毒染色剂用于茎髓内生真菌检测法,包括以下步骤:用解剖刀将禾草成株茎秆纵向剖开,刀尖刮取茎杆髓质放于载玻片上,用染色剂进行染色,盖上盖玻片,并在酒精灯火焰上加热6秒,静置片刻后在显微镜下观察内生真菌菌丝。
以上所述仅为本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (8)
1.一种用于禾草内生真菌检测的无毒染色剂,其特征在于,包括以下成分:苯胺蓝、乳酸和溶剂。
2.根据权利要求1所述的用于禾草内生真菌检测的无毒染色剂,其特征在于,所述的无毒染色剂用于种子内生真菌检测时,溶剂为水。
3.根据权利要求2所述的用于禾草内生真菌检测的无毒染色剂,其特征在于,所述的无毒染色剂用于种子内生真菌检测时,包括以下成分:0.325g苯胺蓝,100ml水,50ml 85%的乳酸。
4.根据权利要求1所述的用于禾草内生真菌检测的无毒染色剂,其特征在于,所述的无毒染色剂用于叶鞘内生真菌检测或茎髓内生真菌检测时,溶剂为丙三醇和水的混合物。
5.根据权利要求4所述的用于禾草内生真菌检测的无毒染色剂,其特征在于,所述的无毒染色剂用于叶鞘内生真菌检测或茎髓内生真菌检测时,包括以下成分:1份乳酸,2份丙三醇,1份蒸馏水,质量分数为0.05%的苯胺蓝。
6.根据权利要求3所述的用于禾草内生真菌检测的无毒染色剂,其特征在于,所述的用于禾草内生真菌检测的无毒染色剂用于种子内生真菌检测法,包括以下步骤:将100粒种子放入20ml烧杯中,倒入5%的氢氧化钠溶液浸泡过夜,自来水冲洗掉碱液,加入染色剂,加热至沸腾再冷却,取出一粒种子放在滴有苯胺蓝无毒染色剂的载玻片上,盖上盖玻片,轻轻按压种子,在物镜10倍和40倍下观察菌丝情况。
7.根据权利要求5所述的用于禾草内生真菌检测的无毒染色剂,其特征在于,所述的用于禾草内生真菌检测的无毒染色剂用于叶鞘内生真菌检测法,包括以下步骤:将带有内生真菌的禾草种子播种于营养土中,取6周龄的幼苗进行带菌率检测,用镊子和解剖刀撕取叶鞘部分内表皮,放于载玻片上加染色剂进行染色,盖上盖玻片,并在酒精灯火焰上加热5-8秒,静置片刻后在显微镜下观察内生真菌菌丝。
8.根据权利要求5所述的用于禾草内生真菌检测的无毒染色剂,其特征在于,所述的用于禾草内生真菌检测的无毒染色剂用于茎髓内生真菌检测法,包括以下步骤:用解剖刀将禾草成株茎秆纵向剖开,刀尖刮取茎杆髓质放于载玻片上,用染色剂进行染色,盖上盖玻片,并在酒精灯火焰上加热5-8秒,静置片刻后在显微镜下观察内生真菌菌丝。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610674349.2A CN106353162A (zh) | 2016-08-16 | 2016-08-16 | 一种用于禾草内生真菌检测的无毒染色剂 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610674349.2A CN106353162A (zh) | 2016-08-16 | 2016-08-16 | 一种用于禾草内生真菌检测的无毒染色剂 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN106353162A true CN106353162A (zh) | 2017-01-25 |
Family
ID=57844896
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610674349.2A Pending CN106353162A (zh) | 2016-08-16 | 2016-08-16 | 一种用于禾草内生真菌检测的无毒染色剂 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106353162A (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109269865A (zh) * | 2018-08-24 | 2019-01-25 | 中南民族大学 | 用于分离摇蚊头壳亚化石的染色剂及其染色方法 |
CN115078043A (zh) * | 2022-07-08 | 2022-09-20 | 珠海美华医疗科技有限公司 | 一种真菌染色液及其制备方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1661357A (zh) * | 2004-02-25 | 2005-08-31 | 兰州大学 | 硬质禾草内生真菌检测技术 |
-
2016
- 2016-08-16 CN CN201610674349.2A patent/CN106353162A/zh active Pending
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1661357A (zh) * | 2004-02-25 | 2005-08-31 | 兰州大学 | 硬质禾草内生真菌检测技术 |
Non-Patent Citations (4)
Title |
---|
李春杰 等: "醉马草内生真菌检测方法的研究", 《2008年中国植病、菌物学会杭州联合年会论文集》 * |
贺冰: "《植物病害组织制片技术》", 30 April 2014, 中国农业出版社 * |
赵晓静: "禾草内生真菌显微结构及其检测方法的研究", 《中国优秀硕士学位论文全文数据库 基础科学辑》 * |
鲍根生 等: "青海高原禾草内生真菌资源的调查和分离", 《草业科学》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109269865A (zh) * | 2018-08-24 | 2019-01-25 | 中南民族大学 | 用于分离摇蚊头壳亚化石的染色剂及其染色方法 |
CN115078043A (zh) * | 2022-07-08 | 2022-09-20 | 珠海美华医疗科技有限公司 | 一种真菌染色液及其制备方法 |
CN115078043B (zh) * | 2022-07-08 | 2024-03-29 | 珠海美华医疗科技有限公司 | 一种真菌染色液及其制备方法 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Ananthanarayan | Ananthanarayan and Paniker's textbook of microbiology | |
Nimrichter et al. | Structure, cellular distribution, antigenicity, and biological functions of Fonsecaea pedrosoi ceramide monohexosides | |
CN106353162A (zh) | 一种用于禾草内生真菌检测的无毒染色剂 | |
Stumpf et al. | Cytological studies of the interactions between the cowpea rust fungus and silicon-depleted French bean plants | |
CN106383047A (zh) | 一种观察植物叶部真菌病害组织病理学过程的染色方法 | |
Dunca et al. | Study of the contaminating microbiota of old paper supports | |
Weil et al. | The Serological Types of Shigella Paradysenteriae (Flexner) I. Types with Single Primary Antigen | |
Ali et al. | Isolation and characterization of a canker disease causing pathogen from Citrus aurantifolia and evaluation of its biological control measure | |
Pomerleau | Pathological anatomy of the Dutch elm disease. Distribution and development of Ceratocystis ulmi in elm tissues | |
Cooley | A study of the physiological relations of Sclerotinia cinerea (Bon.) Schröter | |
Pragathi et al. | Diversity of fungal endophytes and their bioactive metabolites from endemic plants of Tirumala hills-Seshachalam biosphere reserve | |
JPH09275998A (ja) | 菌類の即時判別方法及びその判別装置 | |
Babalola | Pectinolytic and cellulolytic enzymes enhance Fusarium compactum virulence on tubercles infection of Egyptian broomrape | |
CN105548091A (zh) | 小麦光腥黑粉菌的激光共聚焦显微镜检方法 | |
CN102816715B (zh) | 小肠结肠耶尔森菌分离、鉴定的试剂盒、制备和应用 | |
Graham | The basis of the determination of cell viability with the fluorochrome primuline | |
Hutchinson et al. | The effects of fungi on the gametophytes of Pteridium aquilinum (L.) Kühn | |
Onah et al. | Acute oral toxicity studies of the crude extract of endophytic fungi isolated from Annona senegalensis Pers | |
Whitby | Medical Bacteriology, Descriptive and Applied: Including Elementary Helminthology | |
CN107446985A (zh) | 一套苹果果实表层定殖与侵染真菌的快速观察方法 | |
Timothy et al. | Antimicrobial Activities of Chloroform Extract of Whole Plant of Sida acuta Burm. f against Selected Clinical Isolates from Laboratory Samples of a Private Hospital in Benin City, Edo State, Nigeria | |
Kumar | Laboratory manual of microbiology | |
Conn | On the microscopic method of studying bacteria in soil | |
Sălcudean et al. | Partial results regarding the detection and identification of pathogens on dendrofloric plants in different green spaces of Craiova municipality. | |
CN102703566A (zh) | 副溶血性弧菌产毒株鉴定平板试剂盒、制备及使用方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WD01 | Invention patent application deemed withdrawn after publication | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170125 |