JP2013514765A - Cry1BeとCry1Fタンパク質との組合せを用いる昆虫抵抗性管理 - Google Patents
Cry1BeとCry1Fタンパク質との組合せを用いる昆虫抵抗性管理 Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
Description
構造化緩衝帯: トウモロコシ地域の20%の非鱗翅目Btトウモロコシ緩衝帯
ワタ地域の50%の非鱗翅目Bt緩衝帯
ブロック:
内部(すなわち、Bt耕地内)
外部(すなわち、無作為接合を最大化するためのBt耕地の1/2マイル(可能なら1/4マイル)以内の分離された耕地)
圃場内帯状地
帯状地は、幼虫移動の効果を低減するために少なくとも4列の幅(好ましくは6列)でなければならない。」
「コーンボーラーのIRMの要件
−トウモロコシ耕地少なくとも20%に保護ハイブリッド種を植え付ける
−ワタを作る地域では、緩衝帯は50%でなければならない
−保護ハイブリッド種の1/2マイル以内に植え付けるべきである
−緩衝帯は、Bt圃場内に帯状地として植えることができ;緩衝帯の帯状地は少なくとも4列の幅でなければならない
−緩衝帯は、標的昆虫に対して経済的閾値に到達するなら、通常の殺有害生物剤だけで処理することができる
−Btをベースにした噴霧可能な殺虫剤は、緩衝帯のトウモロコシに用いることができない
−適切な緩衝帯は、あらゆる農場にBtトウモロコシを植えるべきである」
Wolfersberger, M.G., (1993), Preparation and Partial Characterization of Amino Acid Transporting Brush Border Membrane Vesicles from the Larval Midgut of the Gypsy Moth (Lymantria Dispar). Arch. Insect Biochem. Physiol. 24: 139-147.
Liang, Y., Patel, S.S., and Dean, D.H., (1995), Irreversible Binding Kinetics of Bacillus thuringiensis Cry1A Delta-Endotoxins to Gypsy Moth Brush Border Membrane Vesicles is Directly Correlated to Toxicity. J. Biol. Chern., 270, 24719-24724.
生物活性
FAW、ECB、ならびにCry1Fa抵抗性FAWおよびECB昆虫に作用する本発明のCryタンパク質のバイオアッセイ結果を表1に示す。双方のタンパク質は、FAW幼虫に対して高度に活性である(この有害生物の考察については、例えば、Tabashnik, PNAS (2008), vol.105, no.49, 19029〜19030を参照されたい)。Cry1Faは、感受性FAWに比較して、Cry1Faの毒性に対して抵抗性であるFAW(rFAW)に対してはるかに活性が低い。Cry1Beは、rFAWに対して、感受性FAWに比較して同じように活性またはより活性である。
結合研究
図1は、スポドプテラ・フルギペルダ(Spodoptera frugiperda)(フォールアーミーワーム、FAW)から産生された刷子縁小胞に対する、125ICry1Fa対Cry1FaまたはCry1Beの競合結合を示す。アッセイは、プルダウン法を使用して二つ組で実施した。FAW−0は、競合リガンドのまったく存在しない状態で受容体に結合された125ICry1Faを表す(対照)。FAW−1,000nM Cry1Faは、その受容体から放射能標識化Cry1Faの結合を置換した相同の非標識化Cry1Faの存在する状態で得られる大きく低下した結合レベルを表す。FAW−1,000nM Cry1Beは、その受容体から放射能標識化Cry1Faの結合を置換できなかった非標識化Cry1Beの存在する状態で得られる結合を表す。
Claims (42)
- Cry1Be殺虫性タンパク質をコードするDNAおよびCry1Fa殺虫性タンパク質をコードするDNAを含むトランスジェニック植物。
- 請求項1に記載の植物の種子。
- Cry1Be殺虫性タンパク質をコードするDNAおよびCry1Fa殺虫性タンパク質をコードするDNAが遺伝子移入されている、請求項1に記載の植物。
- 請求項3に記載の植物の種子。
- 非Bt緩衝帯植物、および請求項1に記載の複数の植物を含む植物の圃場であって、前記緩衝帯植物が、前記圃場の全ての作物植物の40%未満を構成する、圃場。
- 前記緩衝帯植物が、前記圃場の全ての作物植物の30%未満を構成する、請求項5に記載の植物の圃場。
- 前記緩衝帯植物が、前記圃場の全ての作物植物の20%未満を構成する、請求項5に記載の植物の圃場。
- 前記緩衝帯植物が、前記圃場中の全ての作物植物の10%未満を構成する、請求項5に記載の植物の圃場。
- 前記緩衝帯植物が、前記圃場の全ての作物植物の5%未満を構成する、請求項5に記載の植物の圃場。
- 前記緩衝帯植物がブロックまたは帯状地にある、請求項5に記載の植物の圃場。
- 非Bt緩衝帯植物からの緩衝帯種子、および請求項4に記載の複数の種子を含む種子混合物であって、前記緩衝帯種子は混合物の全ての種子の40%未満を構成する、種子混合物。
- 前記緩衝帯種子が、混合物の全ての種子の30%未満を構成する、請求項11に記載の種子混合物。
- 前記緩衝帯種子が、混合物の全ての種子の20%未満を構成する、請求項11に記載の種子混合物。
- 前記緩衝帯種子が、混合物の全ての種子の10%未満を構成する、請求項11に記載の種子混合物。
- 前記緩衝帯種子が、混合物の全ての種子の5%未満を構成する、請求項11に記載の種子混合物。
- 昆虫によるCry毒素に対する抵抗性の発達を管理する方法であって、種子を播いて請求項5に記載の植物の圃場を作製することを含む方法。
- 前記植物が、Cry1Abコア毒素を含むタンパク質をコードするDNAをさらに含む、請求項1に記載のトランスジェニック植物。
- 非Bt緩衝帯植物および請求項17に記載の複数種のトランスジェニック植物を含む植物の圃場であって、前記緩衝帯植物が、前記圃場の全ての作物植物の約20%未満を構成する、圃場。
- 請求項17に記載の複数の植物を含む植物の圃場であって、約10%未満の緩衝帯植物を含む圃場。
- 昆虫によるCry毒素に対する抵抗性の発達を管理する方法であって、種子を播いて請求項19に記載の植物の圃場を作製することを含む方法。
- Cry1Fコア毒素を含むタンパク質およびCry1Beコア毒素を含むタンパク質の双方の有効量を発現する細胞を含む、鱗翅目有害生物を防除するための組成物。
- Cry1Fタンパク質およびCry1Beタンパク質の双方を発現するように形質転換された、微生物または植物細胞である宿主を含む、請求項21に記載の組成物。
- 前記有害生物に対してまたは前記有害生物の環境に対して有効量の請求項21に記載の組成物を提示することを含む、鱗翅目有害生物の防除方法。
- バチルス・チューリンゲンシス(Bacillus thuringiensis)由来の4種の殺虫性タンパク質を産生するトランスジェニック植物であって、前記タンパク質の3種が、第1昆虫に対して非交差反応性活性を示し、かつ前記タンパク質の3種が、第2昆虫に対して非交差反応性活性を示す、トランスジェニック植物。
- 前記昆虫が、ユーロピアンコーンボーラーおよびフォールアーミーワームである、請求項24に記載の植物。
- Cry1Faタンパク質と、Cry1Beタンパク質と、Cry1Ab、Cry2Aa、およびCry1Iタンパク質からなる群から選択される第3タンパク質とを産生するトランスジェニック植物。
- Cry1Faタンパク質と、Cry1Beタンパク質と、Vip3A、Cry1C、Cry1D、およびCry1Eタンパク質からなる群から選択される第3タンパク質とを産生するトランスジェニック植物。
- Cry1Faタンパク質と、Cry1Beタンパク質と、Cry1Ab、Cry2Aa、およびCry1Iタンパク質からなる群から選択される第3タンパク質と、Vip3A、Cry1C、Cry1D、およびCry1Eタンパク質からなる群から選択される第4タンパク質とを産生するトランスジェニック植物。
- 昆虫によるCry毒素に対する抵抗性の発達を管理する方法であって、種子を播いて請求項24から28のいずれかに記載の植物の圃場を作製することを含む方法。
- 非Bt緩衝帯植物、および請求項24から28のいずれかに記載の複数の植物を含む植物の圃場であって、前記緩衝帯植物が、前記圃場の全ての作物植物の約10%未満を構成する、圃場。
- 約5%未満の緩衝帯植物を含む、請求項30に記載の圃場。
- 昆虫によるCry毒素に対する抵抗性の発達を管理する方法であって、種子を播いて請求項30または/および請求項31に記載の植物の圃場を作製することを含む方法。
- 非Bt緩衝帯植物からの緩衝帯種子、および請求項24から28のいずれかに記載の植物からの複数の種子を含む種子混合物であって、前記緩衝帯種子は混合物の全ての種子の10%未満を構成する、種子混合物。
- 前記植物が、10エーカーよりも多くを占める、請求項5、18および30から31のいずれかに記載の圃場。
- 前記植物が、トウモロコシ、ダイズ、およびワタからなる群から選択される、請求項1、2、17、24、25、26および27から28のいずれかに記載の植物。
- 前記植物がトウモロコシ植物である、請求項1、2、17、24、25、26および27から28のいずれかに記載の植物。
- 前記植物細胞が、前記Cry1Fa殺虫性タンパク質をコードする前記DNAおよび前記Cry1Be殺虫性タンパク質をコードする前記DNAを含み、前記Cry1Fa殺虫性タンパク質が配列番号1と少なくとも99%同一であり、前記Cry1Be殺虫性タンパク質が、配列番号2と少なくとも99%同一である、請求項1、2、17、24、25、26および27から28のいずれかに記載の植物の植物細胞。
- 前記Cry1Fa殺虫性タンパク質が配列番号1を含み、前記Cry1Be殺虫性タンパク質が配列番号2を含む、請求項1、2、17、24、25、26および27から28のいずれかに記載の植物。
- 請求項1、2、17、24、25、26および27から28のいずれかに記載の植物の植物細胞であって、前記植物細胞が、前記Cry1Be殺虫性タンパク質をコードする前記DNAおよび前記Cry1Fa殺虫性タンパク質をコードする前記DNAを含み、前記Cry1Be殺虫性タンパク質が、配列番号2と少なくとも99%同一であり、前記Cry1Fa殺虫性タンパク質が、配列番号1と少なくとも99%同一である、植物細胞。
- 前記Cry1Be殺虫性タンパク質が配列番号2を含み、前記Cry1Fa殺虫性タンパク質が配列番号1を含む、請求項1、2、17、24、25、26および27から28のいずれかに記載の植物。
- ユーロピアンコーンボーラーおよびフォールアーミーワームからなる群から選択される昆虫を防除する方法であって、前記昆虫をCry1Be殺虫性タンパク質およびCry1Fa殺虫性タンパク質と接触させることを含む方法。
- 請求項37または請求項39に記載の植物細胞を産生する方法。
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