JP2013521298A - 植物のバイオマスを増やし、鉄濃度を高め、病原体に対する耐性を向上させるための組成物および方法 - Google Patents
植物のバイオマスを増やし、鉄濃度を高め、病原体に対する耐性を向上させるための組成物および方法 Download PDFInfo
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Abstract
Description
本出願は、あらゆる目的のためその全体を本明細書に援用する2010年3月1日に出願された米国特許出願第61/309,134号、2010年11月16日に出願された米国特許出願第61/414,108号、および2010年11月22日に出願された米国特許出願第61/416,039号に対する優先権を主張する。
開示された本発明に至る研究は、University of DelawareのGeorgetown,DEのUSDA Experimental Field Stationからの技術支援と共に、国立科学財団(National Science Foundation:NSF)Grant No.0923806およびNSF Grant No.IOS−0814477により一部資金提供を受けた。したがって、米国政府は、本発明に一定の権利を有する場合がある。
セイヨウヤマカモジ(Brachypodium distachyon)植物およびトウモロコシ植物を発芽させ、21日間生育させた。5日に1回(3回)、ポットごとにOD0.5のB.スブチリス(B.subtilis)FB17 5mlを加えた。対照では、OD0.5の大腸菌(E.coli)OP50 5mlをポットごとに加えた。FB17およびOP50をLB培地で一晩生育させ、SmartSpec(Bio Rad)分光光度計を使用して波長(600nm)の光学密度(OD)を得た。最終処理から10日後、植物を解析した。本明細書のすべての実験に記載される対照とは、細菌で処理しなかったか、または大腸菌(E.coli)OP50で処理した植物をいう。
シロイヌナズナ(Arabidopsis thaliana)の種子を発芽させ、21日間生育させた。5日に1回(3回)、OD0.5のB.スブチリス(B.subtilis)FB17 5mlをポットごとに加えた。対照では、OD0.5の大腸菌(E.coli)OP50 5mlを ポットごとに加えた。FB17およびOP50をLB培地で一晩生育させ、SmartSpec(Bio Rad)分光光度計を使用して波長(600nm)の光学密度(OD)を得た。最終処理から10日後、植物を25℃、湿度40%で4週間乾燥に供した(すなわち、水を加えなかった)。処理から30日後、未処理植物のstay green表現型の喪失をFB17で処理した植物と比較して乾燥を評価したところ、FB17がアラビドプシス属(Arabidopsis)の乾燥耐性を向上させることが示唆された。
B.スブチリス(B.subtilis)FB17の種子処理は、トウモロコシMo17、CML258、CML10、ジニア属(Zinnia)、およびセイヨウヤマカモジ(Brachypodium distachyon)のバイオマスの向上を促進する。
B.スブチリス(B.subtilis)FB17の種子処理は、トウモロコシおよびトマトの光合成効率を促進する。
B.スブチリス(B.subtilis)FB17の種子処理は、トウモロコシ植物およびトマト植物の発芽を促進する。
トウモロコシ(Mo17、CML258、CML10)、ダイズ(Will−82)、トマト(Solanum lycopersicum)、ジニア属(Zinnia)、およびブラキポディウム(Brachypodium)(エネルギー作物モデル)におけるB.スブチリス(B.subtilis)FB17の作用を試験するため、植物種ごとに50の種子(n=50)を、B.スブチリス(B.subtilis)FB17(1e7cfu/種子またはSmartSpec Bio Rad分光光度計を用いて測定した波長600nmの光学密度(OD)0.5の、LB培地で一晩増殖させたバチルス・スブチリス(Bacillus subtilis)FB17 12.5ml/kg)で種子処理した。種子処理後、種子を一つずつポット(4×4インチ)播種した。測定は、処理から15日後に行った。図17は、B.スブチリス(B.subtilis)FB17で処理後のトウモロコシ(Zea mays)の生育速度を図示する。図18は、B.スブチリス(B.subtilis)FB17で処理した植物の保水能力を図示する。FB17処理後、総保水能力および保水率が、トマト(2.1%)およびトウモロコシ(Z.mays)MO17(3.5%)で有意に高まることが観察された。図19は、B.スブチリス(B.subtilis)FB17で処理した植物の乾燥耐性を図示する。MO17の生育速度がFB17処理後、乾燥処理下で有意に高まることが観察された(非水処理対照に対して37.5%増)。図20は、B.スブチリス(B.subtilis)FB17の種子処理がトウモロコシのリグニン含有量を減少させることを図示する。トウモロコシ(Z.mays)ではFB17処理後、総リグニン含有量が有意に減少することが観察された(MO17で約46%減少;CML10で約64%減少、およびCML58で約49%減少)。
図21は、B.スブチリス(B.subtilis)FB17で処理した米植物オリザ・サティバ(Oryza sativa)(日本晴(Nipponbare))の空中および根のバイオマスが接種から60日後に増加することを図示する。LBで一晩生育させたFB17の培養液を用いて108細胞/mlの接種菌液を作製した。FB17の補充には、水耕法で生育させ4週齢の米植物(品種日本晴(Nipponbare))を使用した。B.スブチリス(B.subtilis)FB17を投与した米植物は、未処理米植物と比較して約200%のバイオマスの増加を示した。
バチルス・スブチリス(Bacillus subtilis)FB17が米の根に定着するかどうかを評価するため、米植物(品種日本晴(Nipponbare))にバチルス・スブチリス(Bacillus subtilis)FB17を接種し、接種から96時間後に米植物の根を共焦点走査型レーザー顕微鏡により観察した。観察結果から、この有益な根圏細菌(バチルス・スブチリス(Bacillus subtilis)FB17)は、植物体にバイオフィルムを形成することが確認された。特に、データからは、バチルス・スブチリス(Bacillus subtilis)FB17が処理の96時間後に米の根に効率的に定着することが示唆されることから、米の根が有益な微生物の定着を支持することが示される。
バチルス・スブチリス(Bacillus subtilis)FB17が米の鉄強化を促進するかどうかを評価するため、本出願人らは、誘導結合プラズマ発光分光分析法(ICP−AES)を用いて、バチルス・スブチリス(Bacillus subtilis)FB17を補充した植物の米の葉、根、および穀粒における全鉄含有量を解析した。結果から、バチルス・スブチリス(Bacillus subtilis)FB17を米に補充すると、植物体に鉄が集まりやすくなる、すなわち、必須元素の鉄が、植物の生育および成長に利用される植物に積極的に取り込まれることが明らかになった。図22に図示したように、植物の乾燥重量1kg当たりの鉄のmg数で測定した場合、FB17で処理した米植物では未処理対照と比較して鉄含有量が81%増加することが観察された(図1に示したように「UD1022」とはバチルス・スブチリス(Bacillus subtilis)FB17をいう)。したがって、バチルス・スブチリス(Bacillus subtilis)FB17を植物、特に米植物に投与すると、食物中の鉄濃度の上昇により食物の栄養価を大きく高めることができる。
Claims (29)
- 植物のバイオマスをより多く生産する方法であって、未処理植物と比較して前記植物のバイオマスをより多く生産するのに効果的な量でバチルス・スブチリス(Bacillus subtilis)FB17を前記植物、前記植物の種子、または前記植物もしくは前記種子の周囲の土壌に投与することを含む方法。
- 前記バチルス・スブチリス(Bacillus subtilis)FB17を前記植物の前記種子に投与することを含む、請求項1に記載の方法。
- 前記植物はトウモロコシ植物、ダイズ植物、米植物、およびトマト植物からなる群から選択される、請求項1に記載の方法。
- 前記植物はバイオエネルギー作物植物である、請求項1に記載の方法。
- 前記植物はセイヨウヤマカモジ(Brachypodium distachyon)である、請求項4に記載の方法。
- 未処理植物と比較して前記植物のバイオマスを約5%〜約100%多く生産するのに効果的な量で前記バチルス・スブチリス(Bacillus subtilis)FB17を投与することを含む、請求項1に記載の方法。
- 約1×106CFU/種子〜約1×108CFU/種子の量で前記バチルス・スブチリス(Bacillus subtilis)FB17を前記種子に投与することを含む、請求項2に記載の方法。
- 植物の乾燥耐性を高める方法であって、未処理植物と比較して前記植物の乾燥耐性を高めるのに効果的な量でバチルス・スブチリス(Bacillus subtilis)FB17を前記植物、前記植物の種子、または前記植物もしくは前記種子の周囲の土壌に投与することを含む方法。
- 前記バチルス・スブチリス(Bacillus subtilis)FB17を前記植物の前記種子に投与することを含む、請求項8に記載の方法。
- 前記植物はトウモロコシ植物、ダイズ植物、米植物、およびトマト植物からなる群から選択される、請求項8に記載の方法
- 植物のリグニン濃度を低下させる方法であって、未処理植物と比較して前記植物のリグニン濃度を低下させるのに効果的な量でバチルス・スブチリス(Bacillus subtilis)FB17を前記植物、前記植物の種子、または前記植物もしくは前記種子の周囲の土壌に投与することを含む方法。
- 前記バチルス・スブチリス(Bacillus subtilis)FB17を前記植物の前記種子に投与することを含む、請求項11に記載の方法。
- 前記植物はトウモロコシ植物、ダイズ植物、米植物、およびトマト植物からなる群から選択される、請求項11に記載の方法。
- 約1×106CFU/種子〜約1×108CFU/種子の量で前記バチルス・スブチリス(Bacillus subtilis)FB17を前記種子に投与することを含む、請求項12に記載の方法。
- バイオ燃料を生産する方法であって、バチルス・スブチリス(Bacillus subtilis)FB17が投与された請求項11に記載の植物のバイオマスを前記バイオ燃料に変換することを含む方法。
- 請求項15に従い生産されたバイオ燃料。
- 植物の鉄濃度を上昇させる方法であって、未処理植物と比較して前記植物の鉄濃度を上昇させるのに効果的な量でバチルス・スブチリス(Bacillus subtilis)FB17を前記植物、前記植物の種子、または前記植物もしくは前記種子の周囲の土壌に投与することを含む方法。
- 前記植物は米植物である、請求項17に記載の方法。
- 播く前に前記バチルス・スブチリス(Bacillus subtilis)FB17を前記米植物の前記種子に投与することを含む、請求項18に記載の方法。
- 未処理米植物と比較して前記米植物の鉄濃度を少なくとも約25%上昇させるのに効果的な量で前記バチルス・スブチリス(Bacillus subtilis)FB17を投与することを含む、請求項18に記載の方法。
- 約1×106CFU/種子〜約1×108CFU/種子の量で前記バチルス・スブチリス(Bacillus subtilis)FB17を前記種子に投与することを含む、請求項19に記載の方法。
- 病原真菌による植物の感染を阻害する方法であって、未処理植物と比較して前記病原真菌による前記植物の感染を阻害する量でバチルス・スブチリス(Bacillus subtilis)FB17を前記植物、前記植物の種子、または前記植物もしくは前記種子の周囲の土壌に投与することを含む方法。
- 前記植物は米植物であり、前記病原真菌はイネいもち病である、請求項22に記載の方法。
- 播く前に前記バチルス・スブチリス(Bacillus subtilis)FB17を前記米植物の前記種子に投与することを含む、請求項23に記載の方法。
- 約1×106CFU/種子〜約1×108CFU/種子の量で前記バチルス・スブチリス(Bacillus subtilis)FB17を前記米植物の前記種子に投与することを含む、請求項24に記載の方法。
- 前記イネいもち病の症状は未処理植物と比較して約5%〜約100%軽減される、請求項23に記載の方法。
- バチルス・スブチリス(Bacillus subtilis)FB17を含む農業用担体。
- バチルス・スブチリス(Bacillus subtilis)FB17を含む植物種子コーティング。
- 請求項28に記載のコーティングを含む植物の種子。
Applications Claiming Priority (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US30913410P | 2010-03-01 | 2010-03-01 | |
| US61/309,134 | 2010-03-01 | ||
| US41410810P | 2010-11-16 | 2010-11-16 | |
| US61/414,108 | 2010-11-16 | ||
| US41603910P | 2010-11-22 | 2010-11-22 | |
| US61/416,039 | 2010-11-22 | ||
| PCT/US2011/026683 WO2011109395A2 (en) | 2010-03-01 | 2011-03-01 | Compositions and methods for increasing biomass, iron concentration, and tolerance to pathogens in plants |
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| AU (1) | AU2011223835B2 (ja) |
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| CL (1) | CL2012002419A1 (ja) |
| ES (1) | ES2636649T3 (ja) |
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| JP2021510296A (ja) * | 2018-01-10 | 2021-04-22 | バイエル クロップサイエンス エルピーBayer Cropscience Lp | 改良された微生物とその製造方法 |
| JP2021143188A (ja) * | 2015-08-28 | 2021-09-24 | アグバイオーム, インコーポレイテッド | 植物の病気を制御するための細菌株およびその使用 |
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| AU2011223835B2 (en) | 2010-03-01 | 2015-06-18 | University Of Delaware | Compositions and methods for increasing biomass, iron concentration, and tolerance to pathogens in plants |
| AP2015008329A0 (en) * | 2012-08-31 | 2015-03-31 | Bayer Cropscience Lp | Method of increasing abiotic stress resistance of a plant |
| EP2903438A1 (en) | 2012-10-01 | 2015-08-12 | Basf Se | Pesticidally active mixtures comprising anthranilamide compounds |
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| US20160289130A1 (en) | 2012-11-15 | 2016-10-06 | Basf Corporation | Mulch and Potting Soil Compositions Containing Microorganisms and Related Methods |
| WO2014079820A1 (en) | 2012-11-22 | 2014-05-30 | Basf Se | Use of anthranilamide compounds for reducing insect-vectored viral infections |
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| EP2542047B1 (en) | 2017-05-10 |
| NZ602068A (en) | 2014-11-28 |
| US20110212835A1 (en) | 2011-09-01 |
| PL2542047T3 (pl) | 2018-01-31 |
| US8697603B2 (en) | 2014-04-15 |
| RU2012141561A (ru) | 2014-04-10 |
| WO2011109395A2 (en) | 2011-09-09 |
| AU2011223835B2 (en) | 2015-06-18 |
| CL2012002419A1 (es) | 2014-07-04 |
| US20130184150A1 (en) | 2013-07-18 |
| WO2011109395A3 (en) | 2012-02-02 |
| HUE035280T2 (en) | 2018-05-02 |
| EP2542047A4 (en) | 2013-08-14 |
| CA2791478C (en) | 2019-09-03 |
| BR112012021952A2 (pt) | 2015-09-08 |
| BR112012021952B1 (pt) | 2020-06-09 |
| ES2636649T3 (es) | 2017-10-06 |
| RU2610683C2 (ru) | 2017-02-14 |
| CN103037684A (zh) | 2013-04-10 |
| EP2542047A2 (en) | 2013-01-09 |
| AU2011223835A1 (en) | 2012-09-13 |
| US20140315715A1 (en) | 2014-10-23 |
| MX2012010043A (es) | 2012-12-17 |
| BR112012021952B8 (pt) | 2021-08-17 |
| CA2791478A1 (en) | 2011-09-09 |
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