JP7384347B2 - 微生物凍結乾燥組成物 - Google Patents
微生物凍結乾燥組成物 Download PDFInfo
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- JP7384347B2 JP7384347B2 JP2019551170A JP2019551170A JP7384347B2 JP 7384347 B2 JP7384347 B2 JP 7384347B2 JP 2019551170 A JP2019551170 A JP 2019551170A JP 2019551170 A JP2019551170 A JP 2019551170A JP 7384347 B2 JP7384347 B2 JP 7384347B2
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- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- ZWLUXSQADUDCSB-UHFFFAOYSA-N phthalaldehyde Chemical compound O=CC1=CC=CC=C1C=O ZWLUXSQADUDCSB-UHFFFAOYSA-N 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 235000021018 plums Nutrition 0.000 description 1
- 229920000570 polyether Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920001451 polypropylene glycol Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 229960002816 potassium chloride Drugs 0.000 description 1
- 239000013630 prepared media Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- XOPPYWGGTZVUFP-DLWPFLMGSA-N primeverose Chemical compound O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO[C@@H]1OC[C@@H](O)[C@H](O)[C@H]1O XOPPYWGGTZVUFP-DLWPFLMGSA-N 0.000 description 1
- QYNRIDLOTGRNML-UHFFFAOYSA-N primeverose Natural products OC1C(O)C(O)COC1OCC1C(O)C(O)C(O)C(O)O1 QYNRIDLOTGRNML-UHFFFAOYSA-N 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- HEBKCHPVOIAQTA-ZXFHETKHSA-N ribitol Chemical compound OC[C@H](O)[C@H](O)[C@H](O)CO HEBKCHPVOIAQTA-ZXFHETKHSA-N 0.000 description 1
- OVVGHDNPYGTYIT-BNXXONSGSA-N rutinose Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)O1 OVVGHDNPYGTYIT-BNXXONSGSA-N 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- HSNZZMHEPUFJNZ-SHUUEZRQSA-N sedoheptulose Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)[C@H](O)C(=O)CO HSNZZMHEPUFJNZ-SHUUEZRQSA-N 0.000 description 1
- 230000007226 seed germination Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 229940073490 sodium glutamate Drugs 0.000 description 1
- 235000009518 sodium iodide Nutrition 0.000 description 1
- 244000000000 soil microbiome Species 0.000 description 1
- HIWPGCMGAMJNRG-RTPHMHGBSA-N sophorose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)OC(O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HIWPGCMGAMJNRG-RTPHMHGBSA-N 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000004455 soybean meal Substances 0.000 description 1
- 235000019710 soybean protein Nutrition 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- UQZIYBXSHAGNOE-XNSRJBNMSA-N stachyose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO[C@@H]3[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O3)O)O2)O)O1 UQZIYBXSHAGNOE-XNSRJBNMSA-N 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000000859 sublimation Methods 0.000 description 1
- 230000008022 sublimation Effects 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 150000004044 tetrasaccharides Chemical class 0.000 description 1
- 150000003538 tetroses Chemical class 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 150000003641 trioses Chemical class 0.000 description 1
- 150000004043 trisaccharides Chemical class 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 231100000588 tumorigenic Toxicity 0.000 description 1
- 230000000381 tumorigenic effect Effects 0.000 description 1
- RULSWEULPANCDV-PIXUTMIVSA-N turanose Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](C(=O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RULSWEULPANCDV-PIXUTMIVSA-N 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
- 235000021119 whey protein Nutrition 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 229940102001 zinc bromide Drugs 0.000 description 1
- 239000011592 zinc chloride Substances 0.000 description 1
- 235000005074 zinc chloride Nutrition 0.000 description 1
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
- FYGDTMLNYKFZSV-BYLHFPJWSA-N β-1,4-galactotrioside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-BYLHFPJWSA-N 0.000 description 1
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- C12N1/04—Preserving or maintaining viable microorganisms
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- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
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Description
(1)リゾビウム属菌の生菌を含有してなる凍結乾燥組成物であって、5~10質量%の水分を含有することを特徴とする、微生物凍結乾燥組成物。
(2)リゾビウム属菌が、リゾビウム・ラジオバクター、リゾビウム・リゾゲネス、リゾビウム・ヴィティスから選ばれる1種又は2種以上である、(1)に記載の凍結乾燥組成物。
(3)リゾビウム属菌が非病原性である、(1)又は(2)に記載の凍結乾燥組成物。
(4)リゾビウム属菌が、リゾビウム・ヴィティス ARK-1株(FERM BP-11426)、リゾビウム・ヴィティス ARK-2株(FERM BP-11427)、リゾビウム・ヴィティス ARK-3株(FERM BP-11428)、アグロバクテリウム・ラジオバクター K84株から選ばれる1種又は2種以上である、(3)に記載の凍結乾燥組成物。
(5)(1)~(4)のいずれか1つに記載の凍結乾燥組成物を0~10℃で保存することを特徴とする、凍結乾燥組成物中のリゾビウム属菌生菌生存率低下抑制方法。
(6)(3)又は(4)に記載の凍結乾燥組成物を水に投入して分散させた非病原性リゾビウム属菌の生菌の懸濁液に、定植前の有用植物の苗木を浸漬することを特徴とする、根頭がんしゅ病の防除方法。
(7)(3)又は(4)に記載の凍結乾燥組成物を微粉砕し、該粉砕物を含む非病原性リゾビウム属菌の生菌を含有する粉体を、定植前の有用植物の苗木に粉衣することを特徴とする、根頭がんしゅ病の防除方法。
(8)リゾビウム属菌の生菌と10質量%を超える水分を含有してなる組成物を凍結乾燥する工程を含み、前記工程は、該組成物を凍結後に水分含量が10質量%となるまで減圧乾燥して以降、水分含量が5質量%未満となる以前に該乾燥を終了して、リゾビウム属菌の生菌と5~10質量%の水分を含有する凍結乾燥組成物を得る工程であることを特徴とする、微生物凍結乾燥組成物の製造方法。
(9)リゾビウム属菌が、リゾビウム・ラジオバクター、リゾビウム・リゾゲネス、リゾビウム・ヴィティスから選ばれる1種又は2種以上である、(8)に記載の方法。
(10)リゾビウム属菌が非病原性である、(8)又は(9)に記載の方法。
(11)リゾビウム属菌が、リゾビウム・ヴィティス ARK-1株(FERM BP-11426)、リゾビウム・ヴィティス ARK-2株(FERM BP-11427)、リゾビウム・ヴィティス ARK-3株(FERM BP-11428)、アグロバクテリウム・ラジオバクター K84株から選ばれる1種又は2種以上である、(10)に記載の方法。
<NA(Nutrient Agar)平板培地の調製>
肉エキス5部、ペプトン10部、塩化ナトリウム5部、寒天15部及び水1000部を混合し、加熱して完全に溶解させた後、シャーレに流し入れ、冷却して、NA平板培地を調製した。
大豆ペプチド17部、酵母エキス3部、グルコース2.5部、塩化ナトリウム5部及び水1000部を混合して前培養用培地を調製した。
グルコース2.5部をマルトース20部に変更し、消泡剤としてポリオキシアルキレングリコール5部を添加したこと以外は前培養用培地の調製と同様にして、本培養用培地を調製した。
トレハロース20部、グルタミン酸ナトリウム10部、システイン塩酸塩8部カルボキシメチルセルロースナトリウム1部及び水61部を混合し、若干量の水酸化ナトリウム水溶液でpH7に調整して保護剤水溶液を調製した。
リゾビウム・ヴィティス ARK-1株のグリセロールストックを解凍して前記NA平板培地に植菌し、28℃で1日間静置培養した。形成したコロニーを白金耳にて1エーゼ掻き取り、前記前培養用培地に植菌し、26時間撹拌培養した。この水性培養液を前記本培養用培地に植菌し、28℃で43時間撹拌培養した。本培養終了後の水性培養液を遠心分離して上澄みを捨て、リゾビウム・ヴィティス ARK-1株の生菌の濃縮物を得た。該濃縮物50部と前記保護剤水溶液50部を混合し、若干量の水酸化ナトリウム水溶液を滴下してpH7に調整し、生菌原液を調製した。生菌原液中のリゾビウム・ヴィティス ARK-1株の生菌数濃度を前記NA平板培地を用いて希釈平板法により測定したところ3.6×1011CFU/mLであった。
[実施例1]
前記生菌原液25gを100mL容ナスフラスコに取り、-80℃に設定したディープフリーザーで予備凍結した後、凍結乾燥装置(FDU-2100型、東京理化器械株式会社製品)を用いて、ナスフラスコ内を5~10Pa程度に減圧して凍結乾燥を15時間実施し、凍結乾燥組成物を得た。ナスフラスコ内の凍結乾燥組成物の全質量は8.1g、水分含量は6.3質量%であった。
凍結乾燥を6時間としたこと以外は実施例1と同様にして、凍結乾燥組成物を得た。ナスフラスコ内の凍結乾燥組成物の全質量は11.6g、水分含量は25.9質量%であった。この凍結乾燥組成物は粘着性を有した。
凍結乾燥を10時間としたこと以外は実施例1と同様にして、凍結乾燥組成物を得た。ナスフラスコ内の凍結乾燥組成物の全質量は9.5g、水分含量は14.8質量%であった。この凍結乾燥組成物は粘着性を有した。
凍結乾燥を24時間としたこと以外は実施例1と同様にして、凍結乾燥組成物を得た。ナスフラスコ内の凍結乾燥組成物の全質量は7.1g、水分含量は1.5質量%であった。
凍結乾燥を48時間としたこと以外は実施例1と同様にして、凍結乾燥組成物を得た。ナスフラスコ内の凍結乾燥組成物の全質量は7.0g、水分含量は0.6質量%であった。
前期実施例1及び比較例1~4の凍結乾燥組成物中のリゾビウム・ヴィティス ARK-1株の生菌数濃度(CFU/g)を、前記NA平板培地を用いて希釈平板法により測定し、これに調製された凍結乾燥組成物の全質量(g)を乗じて、凍結乾燥組成物中のリゾビウム・ヴィティス ARK-1株総生菌数(CFU)を求めた。この総生菌数を前記生菌原液25g中のリゾビウム・ヴィティス ARK-1株の総生菌数9×1012CFU(=3.6×1011CFU/g×25g)で除し、凍結乾燥後のリゾビウム・ヴィティス ARK-1株の生菌生存率(%)を算出した。結果を表1に示す。
<生菌原液の調製>
リゾビウム・ヴィティス ARK-1株のグリセロールストックを解凍して前記NA平板培地に植菌し、28℃で1日間静置培養した。形成したコロニーを白金耳にて1エーゼ掻き取り、前培養用培地に植菌し、25時間撹拌培養した。この水性培養液を前記本培養用培地に植菌し、30Lジャーファーメンターを用いて28℃で42時間撹拌培養した。本培養終了後の水性培養液を遠心分離して上澄みを捨て、リゾビウム・ヴィティス ARK-1株の生菌の濃縮物を得た。該濃縮物50部と試験例1で作製した保護剤水溶液50部を混合し、若干量の水酸化ナトリウム水溶液を滴下してpH7に調整し、生菌原液を調製した。生菌原液中のリゾビウム・ヴィティス ARK-1株の生菌数濃度を前記NA平板培地を用いて希釈平板法により測定したところ3.5×1011CFU/mLであった。
前記生菌原液7.0gを10mL容バイアル瓶に取り、-80℃に設定したディープフリーザーで予備凍結した後、凍結乾燥装置(Vir Tis25L Genesis SQ Super ES-55、SP Industries,Inc.製品)を用いて、200m Torr程度に減圧して凍結乾燥を48時間実施し、凍結乾燥組成物を得た。バイアル瓶内の凍結乾燥組成物の全質量は2.0g、水分含量は7.0質量%であった。
前記凍結乾燥組成物を密封したバイアル瓶のまま4℃の冷蔵室又は28℃の恒温室で静置保存した。所定期間保存した後、バイアル瓶内のリゾビウム・ヴィティス ARK-1株の生菌数濃度(CFU/g)を、前記NA平板培地を用いて希釈平板法により測定し、これに調製された凍結乾燥組成物の全質量(g)を乗じて、凍結乾燥組成物中のリゾビウム・ヴィティス ARK-1株総生菌数(CFU)を求めた。この総生菌数を前記生菌原液7.0g中のリゾビウム・ヴィティス ARK-1株の総生菌数2.5×1012CFU(=3.5×1011CFU/g×7.0g)で除し、保存後のリゾビウム・ヴィティス ARK-1株の生菌生存率(%)を算出した。4℃保存の結果を表2、28℃保存の結果を表3に示す。
NA(Nutrient Agar)平板培地の調製、前培養用培地の調製、及び保護剤水溶液の調製は、試験例1と同様にして行い、本培養用培地の調製は次のようにして行った。
大豆ペプチド102部、酵母エキス18部、塩化ナトリウム30部、マルトース120部、ディスホームNQH-7403(消泡剤)を30部及び水6000部を混合して調製した。
リゾビウム・ヴィティス ARK-1株、リゾビウム・ヴィティス ARK-2株、リゾビウム・ヴィティス ARK-3株及びアグロバクテリウム・ラジオバクター K84株のグリセリンストックを解凍して前記NA平板培地に植菌し、28℃で1日間静置培養した。前記前培養培地に種菌より一白金耳量を接種し、28℃の恒温室内で190rpmのロータリーシェーカーを用いて1日間前培養を行った。前培養液をそれぞれ接種して、10Lジャーファーメンターを用いて28℃で2日間本培養を行った。本培養中は、リゾビウム・ヴィティス ARK-1株、リゾビウム・ヴィティス ARK-2株及びリゾビウム・ヴィティス ARK-3株はpHが7.0を超えないように、リン酸を用いてpH7.0以下制御とし、アグロバクテリウム・ラジオバクター K84株はpH6.0を超えないようにリン酸を用いてpH6.0以下制御で本培養を行った。本培養終了後の培養液を遠心分離して上澄みを捨て、生菌の濃縮物を得た。該濃縮物50部と前記保護剤水溶液50部を混合し、若干量の水酸化ナトリウム水溶液を滴下してpH7に調整し、生菌原液を調製した。生菌原液中の生菌数濃度を常法により測定した。
前記各菌株の生菌原液25gを100mL容ナスフラスコに取り、-80℃に設定したディープフリーザーで予備凍結した後、凍結乾燥装置(FDUー2100型、東京理化器械株式会社製品)を用いて、ナスフラスコ内を5~10Pa程度に減圧して、凍結時間を変えて凍結乾燥を実施し、凍結乾燥組成物を得た。
前記各凍結乾燥組成物中の菌株の生菌数濃度(cfu/g)を常法により測定し、これに調製された凍結乾燥組成物の全質量(g)を乗じて、凍結乾燥組成物中の総生菌数(cfu)を求めた。この総生菌数を前記各生菌原液25g中の菌株の総生菌数で除し、凍結乾燥後の各菌株の生菌生存率(%)を算出した。
(1)リゾビウム・ヴィティス(Rhizobium vitis)ARK-1株(FERM BP-11426)
(2)リゾビウム・ヴィティス(Rhizobium vitis)ARK-2株(FERM BP-11427)
(3)リゾビウム・ヴィティス(Rhizobium vitis)ARK-3株(FERM BP-11428)
Claims (4)
- リゾビウム(Rhizobium)属菌の生菌と5~10質量%の水分を含有してなる微生物凍結乾燥組成物の製造方法であって、該リゾビウム(Rhizobium)属菌が、リゾビウム・ラジオバクター(Rhizobium radiobacter)、リゾビウム・リゾゲネス(Rhizobium rhizogenes)、リゾビウム・ヴィティス(Rhizobium vitis)から選ばれる1種又は2種以上であり、該生菌と10質量%を超える水分を含有してなる組成物を凍結乾燥する工程を含み、前記工程は、該組成物を凍結後に水分含量が10質量%となるまで減圧乾燥して以降、水分含量が5質量%未満となる以前に該乾燥を終了して、該生菌と5~10質量%の水分を含有する凍結乾燥組成物を得る工程である、微生物凍結乾燥組成物の製造方法により製造した微生物凍結乾燥組成物を0~10℃で保存することを特徴とする、該微生物凍結乾燥組成物中のリゾビウム(Rhizobium)属菌生菌生存率低下抑制方法。
- リゾビウム(Rhizobium)属菌の生菌と5~10質量%の水分を含有してなる微生物凍結乾燥組成物の製造方法であって、該リゾビウム(Rhizobium)属菌が、リゾビウム・ヴィティス(Rhizobium vitis)ARK-1株(FERM BP-11426)、リゾビウム・ヴィティス(Rhizobium vitis)ARK-2株(FERM BP-11427)、リゾビウム・ヴィティス(Rhizobium vitis)ARK-3株(FERM BP-11428)、アグロバクテリウム・ラジオバクター(Agrobacterium radiobacter)K84株から選ばれる1種又は2種以上であり、該生菌と10質量%を超える水分を含有してなる組成物を凍結乾燥する工程を含み、前記工程は、該組成物を凍結後に水分含量が10質量%となるまで減圧乾燥して以降、水分含量が5質量%未満となる以前に該乾燥を終了して、該生菌と5~10質量%の水分を含有する凍結乾燥組成物を得る工程である、微生物凍結乾燥組成物の製造方法により製造した微生物凍結乾燥組成物を0~10℃で保存することを特徴とする、該微生物凍結乾燥組成物中のリゾビウム(Rhizobium)属菌生菌生存率低下抑制方法。
- リゾビウム(Rhizobium)属菌の生菌と5~10質量%の水分を含有してなる微生物凍結乾燥組成物の製造方法であって、該リゾビウム(Rhizobium)属菌が、リゾビウム・ヴィティス(Rhizobium vitis)ARK-1株(FERM BP-11426)、リゾビウム・ヴィティス(Rhizobium vitis)ARK-2株(FERM BP-11427)、リゾビウム・ヴィティス(Rhizobium vitis)ARK-3株(FERM BP-11428)、アグロバクテリウム・ラジオバクター(Agrobacterium radiobacter)K84株から選ばれる1種又は2種以上であり、該生菌と10質量%を超える水分を含有してなる組成物を凍結乾燥する工程を含み、前記工程は、該組成物を凍結後に水分含量が10質量%となるまで減圧乾燥して以降、水分含量が5質量%未満となる以前に該乾燥を終了して、該生菌と5~10質量%の水分を含有する凍結乾燥組成物を得る工程である、微生物凍結乾燥組成物の製造方法により製造した微生物凍結乾燥組成物を水に投入して、リゾビウム(Rhizobium)属菌の生菌の懸濁液を得た後、定植前の有用植物の苗木を当該懸濁液に浸漬することを特徴とする、根頭がんしゅ病の防除方法。
- リゾビウム(Rhizobium)属菌の生菌と5~10質量%の水分を含有してなる微生物凍結乾燥組成物の製造方法であって、該リゾビウム(Rhizobium)属菌が、リゾビウム・ヴィティス(Rhizobium vitis)ARK-1株(FERM BP-11426)、リゾビウム・ヴィティス(Rhizobium vitis)ARK-2株(FERM BP-11427)、リゾビウム・ヴィティス(Rhizobium vitis)ARK-3株(FERM BP-11428)、アグロバクテリウム・ラジオバクター(Agrobacterium radiobacter)K84株から選ばれる1種又は2種以上であり、該生菌と10質量%を超える水分を含有してなる組成物を凍結乾燥する工程を含み、前記工程は、該組成物を凍結後に水分含量が10質量%となるまで減圧乾燥して以降、水分含量が5質量%未満となる以前に該乾燥を終了して、該生菌と5~10質量%の水分を含有する凍結乾燥組成物を得る工程である、微生物凍結乾燥組成物の製造方法により製造した微生物凍結乾燥組成物を微粉砕し、該粉砕物を含むリゾビウム(Rhizobium)属菌の生菌を含有する粉体を、定植前の有用植物の苗木に粉衣することを特徴とする、根頭がんしゅ病の防除方法。
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