IP Library › Granted Patent US 10,294,490

Granted Patent B2

US 10,294,490 · App. 15/274,758 · Granted May 21, 2019

Insecticidal proteins and methods for their use

Inventors: Daniel J Altier (Granger, IA); Jennifer K Barry (Ames, IA); Carol A Hendrick (Des Moines, IA); Lu Liu (Palo Alto, CA); Phillip A Patten (Portola Valley, CA); Claudia D Perez-Ortega (Wilmington, DE); Eric J Schepers (Port Deposit, MD); Weiping Xie (East Palo Alto, CA); Nasser Yalpani (Johnston, IA); Jianzhou Zhao (Johnston, IA); Xiaohong Zhong (San Leandro, CA); Genhai Zhu (San Jose, CA)

Assignees: PIONEER HI-BRED INTERNATIONAL, INC.; E I DU PONT NEMOURS AND COMPANY

C12N15/8286 A01N37/46 A01N63/02 C07K14/195 C07K14/225 C12N15/8205 C12N15/8207 C07K2319/00 C07K2319/21 Y02A40/162

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Quick Facts

Patent No.: US 10,294,490
App. No.: 15/274,758
Granted: May 21, 2019
Kind: B2

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Abstract

Compositions and methods for controlling pests are provided. The methods involve transforming organisms with a nucleic acid sequence encoding an insecticidal protein. In particular, the nucleic acid sequences are useful for preparing plants and microorganisms that possess insecticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions are insecticidal nucleic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors for subsequent transformation into organisms of interest including plants, as probes for the isolation of other homologous (or partially homologous) genes. The pesticidal proteins find use in controlling, inhibiting growth or killing Lepidopteran, Coleopteran, Dipteran, fungal, Hemipteran and nematode pest populations and for producing compositions with insecticidal activity.

Claims (593)

1. A recombinant AflP-1A polypeptide having at least 85% identity to the amino acid sequence of SEQ ID NO: 2, and comprising the amino acid motifs as represented by positions 15-26 of SEQ ID NO: 257, positions 33-53 of SEQ. ID NO: 257, positions 71-84 of SEQ ID NO: 257, and positions 100-107 of SEQ ID NO: 257.

2. The recombinant AfIP-1A polypeptide of claim 1 , wherein the AfIP-1A polypeptide comprises an amino acid sequence of the formula

(SEQ ID NO: 257)

1 5 10 15

Met Thr Ala Lys Asp Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala

20 25 30

Xaa Xaa Gln Xaa Xaa Glu Ile Xaa Xaa Phe Xaa Xaa Asn Xaa Xaa

35 40 45

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

50 55 60

Xaa Xaa Xaa Lys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa

65 70 75

Xaa Xaa Ile Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ile Ala Xaa Xaa Xaa

80 85 90

Xaa Xaa Xaa Xaa Ser Ser Xaa Thr Glu Gly Xaa Phe Xaa Xaa Xaa

95 100 105

Xaa Xaa Asp Lys Xaa Xaa Xaa Xaa Tyr Xaa Xaa Asp Cys Xaa Xaa

110 115 120

Xaa Gly Ser Asn Ser Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

125 130 135

Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

140 145

Xaa Xaa Xaa Xaa Xaa Thr Xaa Xaa Lys Xaa Xaa,

wherein

Xaa at position 6 is Ile, Thr, Leu, Met, Val or Ser;

Xaa at position 7 is Ala, Met, Val, Leu, Ile or Gly;

Xaa at position 8 is Thr, Asp, Ser or Glu;

Xaa at position 9 is Glu, Leu, Gly, Asp, Ala, Ile, Val or Met;

Xaa at position 10 is Glu, Asn, Asp or Gln;

Xaa at position 11 is Ser, Val, Thr, Ile, Leu or Met;

Xaa at position 12 is Lys, Glu, Arg or Asp;

Xaa at position 13 is Ile, Val, Leu or Met;

Xaa at position 14 is Arg, Gln, Lys or Asn;

Xaa at position 16 is Tyr, Gln, Trp, Phe or Asn;

Xaa at position 17 is Ala, Ser, Gly or Thr;

Xaa at position 19 is Trp, Glu, Phe, Ile, His, Asn or Tyr;

Xaa at position 20 is Ile, Val, Ala, Cys, Glu, Phe, Gly, Met, Asn, Gln, Arg, Ser or Thr;

Xaa at position 23 is Thr, Glu, Ala, Ser, Asp or Gly;

Xaa at position 24 is Ile, Leu, Val or Met;

Xaa at position 26 is Val, Ser, Ile, Leu, Met or Thr;

Xaa at position 27 is Val, Glu, Ile, Leu, Met or Asp;

Xaa at position 29 is Ser, Met, Thr, Ile, Leu or Val;

Xaa at position 30 is Asn, Asp, Ser, Glu, Gln or Thr;

Xaa at position 31 is Phe, Ile, Leu, Val or Met;

Xaa at position 32 is Lys, Glu, Arg or Asp;

Xaa at position 33 is Val, Ile, Leu or Met;

Xaa at position 34 is Glu, Lys, Asp or Arg;

Xaa at position 35 is Gly or Asn;

Xaa at position 36 is Ala, Gly, Asp, Glu, Phe, Gly, Ile, Leu, Pro, Gln, Arg, Ser, Thr, Val, Trp or Tyr;

Xaa at position 37 is Tyr, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Pro, Arg, Ser, Thr, Val or Trp;

Xaa at position 38 is Leu, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Gln, Arg, Ser, Thr, Val, Trp or Tyr;

Xaa at position 39 is Arg, Lys, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Asn, Pro, Ser, Thr, Val, Trp or Tyr;

Xaa at position 40 is Trp, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val or Tyr;

Xaa at position 41 is Gly, Cys or Gln;

Xaa at position 42 is Lys, Cys, Glu, His, Leu, Met, Asn, Gln, Arg or Thr;

Xaa at position 43 is Phe, Tyr, Ala, Cys, Glu, Ile, Leu, Met, Gln, Ser, Val or Trp;

Xaa at position 44 is His, Ala, Asp, Glu, Gly, Lys, Leu, Met, Asn, Pro, Glu, Arg, Ser, Thr, Val, Trp;

Xaa at position 45 is Val, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Ser, Thr or Trp;

Xaa at position 46 is Pro, Ala, Cys, Asp, Glu, Gly, His, Lys, Leu, Met, Gln, Arg, Ser, Thr, Val, Trp or Tyr;

Xaa at position 47 is Gly, Leu or Phe;

Xaa at position 48 is Asp, Asn, Glu, Gln, Leu or Phe;

Xaa at position 50 is Asp, Ser, Glu, Thr, Leu or Phe;

Xaa at position 51 is Lys, Asn, Arg, Gln, Leu or Phe;

Xaa at position 52 is Glu, Leu or Phe;

Xaa at position 53 is Ile, Leu or Phe;

Xaa at position 54 is Ser, Thr, Leu or Phe;

Xaa at position 55 is Pro, Ser, Thr, Leu or Phe;

Xaa at position 56 is Ser, Asp, Thr, Glu, Leu;

Xaa at position 57 is Gln, Thr, Glu, Asn, Ser, Asp, Leu or Phe;

Xaa at position 58 is Ile, Val, Leu, Met or Phe;

Xaa at position 60 is Gly, Lys, Ala or Arg;

Xaa at position 61 is Thr, Ile or Phe;

Xaa at position 62 is Ile, Lys, Val, Leu, Met, Arg or Phe;

Xaa at position 64 is Lys, Ser, Glu, Arg, Thr, Asp, Leu or Phe;

Xaa at position 65 is Asp, Ser, Glu, Thr, Leu or Phe;

Xaa at position 66 is Glu, Gly, Asp or Ala;

Xaa at position 67 is Asp, Thr, Glu or Ser;

Xaa at position 68 is Ser, Lys, Thr or Arg;

Xaa at position 69 is Tyr, Ser, Trp, Phe or Thr;

Xaa at position 70 is Thr, Lys, Ser or Arg;

Xaa at position 73 is Ser, Ala, Thr or Gly;

Xaa at position 74 is Cys, Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Ser, Thr or Tyr;

Xaa at position 76 is Arg, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Ser, Thr, Val, Trp or Tyr;

Xaa at position 77 is Glu, Ala, Asp or Gly;

Xaa at position 78 is Asn, Asp, Gln or Glu;

Xaa at position 79 is Ala, Thr, Gly or Ser;

Xaa at position 82 is Gly, Glu, Asn, Trp or Tyr;

Xaa at position 86 is Gly, Lys, Ala or Arg;

Xaa at position 88 is Ser, Glu, Thr or Asp;

Xaa at position 89 is Leu, Ile, Val or Met;

Xaa at position 91 is Asp, His or Glu;

Xaa at position 92 is Gly, Asp, Ala or Glu;

Xaa at position 95 is Leu, Trp, Ile, Val, Met, Phe or Tyr;

Xaa at position 96 is Val, Leu, Ile or Met;

Xaa at position 97 is Phe, Ala or Gly;

Xaa at position 98 is Glu, Thr, Asp or Ser;

Xaa at position 100 is Tyr, Lys, Trp or Arg;

Xaa at position 101 is Trp, Phe or Tyr;

Xaa at position 104 is Pro, Ala, Phe, Gly, His, Met, Gln, Arg or Val;

Xaa at position 105 is Trp, Asp, Phe, Ile, Leu or Tyr;

Xaa at position 106 is Ser, Ala, Thr or Gly;

Xaa at position 111 is Asp, His, Asn, Glu or Gln;

Xaa at position 112 is Glu, Ser, Asp or Thr;

Xaa at position 113 is Leu, Ser, Ile, Val, Met or Thr;

Xaa at position 114 is Thr or Ser;

Xaa at position 115 is Val, Ile, Val or Met;

Xaa at position 116 is Lys, Thr, Glu, Arg, Ser or Asp;

Xaa at position 117 is Asp or Glu;

Xaa at position 118 is Lys, Asp, Arg or Glu;

Xaa at position 119 is Glu, Asn, Asp or Gln;

Xaa at position 120 is Asn, Lys, Asp or Arg;

Xaa at position 121 is Tyr, Leu or Phe;

Xaa at position 122 is Thr, Lys, Ser, Arg, Leu or Phe;

Xaa at position 123 is Val, Ile, Leu, Met, Phe or Asn;

Xaa at position 124 is Ile, Ser, Asp, Leu, Val Met, Thr, Glu or Phe;

Xaa at position 125 is Lys, Leu, Phe or Met;

Xaa at position 126 is Lys, Glu, Arg, Asp, Leu or Phe;

Xaa at position 128 is Gly, Leu or Phe;

Xaa at position 129 is Gly, Asn, Ala, Gln, Leu or Phe;

Xaa at position 130 is Ser, Ile, Thr, Leu, Val, Met or Phe;

Xaa at position 131 is Pro, Ser or Thr;

Xaa at position 132 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp or Tyr;

Xaa at position 133 is Gly, Ala, Cys, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val or Trp;

Xaa at position 134 is Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp or Tyr;

Xaa at position 135 is Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Val, Trp or Tyr;

Xaa at position 136 is Gly, Ala, Cys, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp or Tyr;

Xaa at position 137 is Asn, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Glu, Arg, Ser, Thr, Val, Trp or Tyr;

Xaa at position 138 is Ile, Ala, Cys, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp or Tyr;

Xaa at position 139 is Phe, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp or Tyr;

Xaa at position 140 is Ile, Ala, Cys, Phe, His, Leu, Met, Asn, Gln, Thr, Val or Tyr;

Xaa at position 142 is Val, Cys, Ile, Leu or Met;

Xaa at position 143 is Val, Ile; Leu or Met;

Xaa at position 145 is Lys, Val, Arg, Ile, Leu or Met; and

Xaa at position 146 is Ser, Gly, Thr or Ala;

and wherein, 1 to 14 amino acids are optionally deleted from the N-terminus of the polypeptide.

3. The recombinant AfIP-1A polypeptide of claim 1 , wherein the AfIP-1A polypeptide comprises the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 94.

4. A composition, comprising a pesticidally-effective amount of the recombinant AfIP-1A polypeptide of claim 1 .

5. A recombinant AfIP-1B polypeptide having at least 95% identity to the amino acid sequence of SEQ ID NO: 4.

6. The recombinant AfIP-1B polypeptide of claim 5 , comprising the amino acid motifs as represented by positions 105-115 of SEQ ID NO: 259, positions 133-144 of SEQ ID NO: 259, positions 177-184 of SEQ ID NO: 259, positions 358-365 of SEQ ID NO: 259, and positions 511-520 of SEQ ID NO: 259.

7. The recombinant AfIP-1B polypeptide of claim 5 , wherein the AfIP-1 B polypeptide comprises an amino acid sequence of the formula

SEQ ID NO: 259

Met Asp Ile Glu Ala Lys Ser Ile Asn Pro Leu Xaa Gly Ile Thr Val

1 5 10 15

Gly Ser Thr Val Leu Gly Asn Asp Phe Lys Pro Ser Val Leu Ile Asn

20 25 30

Pro Xaa Thr Arg Met Xaa Thr Gln Lys Xaa Xaa Glu Met Gly Met Ser

35 40 45

Gly Lys Ser Gln Xaa Val Xaa Thr Asp Ser Leu Asn Glu Gly Thr Ile

50 55 60

Gly Phe Ser Gly Ala Tyr Xaa Pro Ser Gly Ile Ala Lys Phe Thr Ser

65 70 75 80

Ala Val Ala Val Ser Xaa Ser Asn Ala Thr Ala Ser Glu Xaa Lys Ser

85 90 95

Xaa Lys Val Ser Xaa Asn Xaa Ser Xaa Xaa Ser Xaa Xaa Xaa Xaa Ile

100 105 110

Asp Phe Xaa Asn Leu Thr Val Glu Asp Val Leu Asn Ser Leu Ser Ala

115 120 125

Gly Pro Lys Asn Leu Xaa Leu Lys Xaa Leu Xaa Lys Xaa Ile Ala Xaa

130 135 140

Arg Asp Cys Xaa Gly Ser Ser Xaa Glu Lys Xaa Glu Leu Met Lys Glu

145 150 155 160

Trp Val Lys Ser Leu Gln Asn Phe Ile Ser Ser Tyr Gly Asp Gly Leu

165 170 175

Val Val Xaa Ala Xaa Xaa Gly Gly Met Gly Ser Xaa Ser Thr Glu Met

180 185 190

Thr Ser Lys Xaa Xaa Glu Asp Ser Xaa Xaa Xaa Gly Glu Thr Ala Xaa

195 200 205

Phe Ser Tyr Ser Gly Xaa Gly Ser Ser Val Ser Xaa Ala Gln Thr Xaa

210 215 220

Asn Gly Ser Gln Lys Asp Gln Ser Ser Xaa Xaa Glu Val Ser Cys Lys

225 230 235 240

Ala Leu Ala Ser Gly Gly Cys Val Glu Ser Gln Val Asn Ser Trp Phe

245 250 255

Asp Val Val Ala Asn Lys Ser Phe Ala Glu Ile Ser Gly Xaa Ser Leu

260 265 270

Leu Asp Lys Ala Pro Met Gln Ser Ser Val Ser Pro Pro Pro Lys Ile

275 280 285

Pro Asp Phe Leu Lys Pro Glu Xaa Asn Xaa Xaa Xaa Thr Glu Lys Leu

290 295 300

Xaa Thr Ile Lys Lys Leu Gly Asp Ser Glu Glu Phe Xaa Leu Ala Ser

305 310 315 320

Gly Tyr Xaa Glu Ala Lys Lys Thr Asn Pro Asn Leu Thr Phe Xaa Glu

325 330 335

Phe Lys Ser Thr Val Arg Asp Lys Asn Asn Ile Asp Gly Leu Asn Xaa

340 345 350

Leu Ala Ser Lys Val Gln Xaa Xaa Xaa Leu Xaa Xaa Xaa Ala Xaa Gly

355 360 365

Ser Xaa Ser Arg Xaa Xaa Lys Asn Xaa Ser Leu Gln Gly Val Arg Xaa

370 375 380

Xaa Ser Leu Xaa Ser Ser Ser Asp Xaa Ala Val Leu Gly Xaa Trp Ile

385 390 395 400

Ala Asn Trp Ser Asp Ile Phe Pro Trp Met Ser Met Gly Xaa Met Asn

405 410 415

Glu Xaa Xaa Asp Ala Glu Xaa Ala Xaa Tyr Xaa Leu Lys Ile Arg Cys

420 425 430

Met Xaa Gln Asp Leu Ser Thr Leu Asn Thr Ile Tyr Asn Thr Phe Asn

435 440 445

Ala Cys Asn Ile Lys Leu Asp Phe Cys His Leu Asn Ser Ala Ser Gln

450 455 460

Val Ala Asp Ser Phe Lys Ile Ala Gln Gly Val Leu Ser Asp Asn Val

465 470 475 480

Xaa Ser Asp Asp Ala Val Glu Ile Ala Phe Asn Ser Leu Ser Xaa Glu

485 490 495

Ala Lys Lys Ile Tyr Thr Ala Trp Asn Glu Ile Gly Xaa Leu Arg Xaa

500 505 510

Ala Xaa Leu Xaa Leu Gly Xaa Leu Ile Gly Asp Gln Ser Xaa Ser Ser

515 520 525

Glu Xaa Ile Asp Xaa Lys Pro Xaa Pro Xaa Pro Glu Val Thr Xaa Xaa

530 535 540

Ala Ala Xaa Cys Ser Xaa Gly Xaa Asn Asn Pro Thr Ala Xaa Ser Ser

545 550 555 560

Phe Ile Lys Met Leu Pro Phe Ile Asp Thr Asn Gly Asp Ile Tyr Ala

565 570 575

Phe Gly Pro Ser Leu Met Leu Leu Arg Lys Ala Leu Pro Glu Lys Met

580 585 590

Ile Phe Thr Lys Gly Gly Glu Xaa Ala Xaa Lys Leu Thr Ala Xaa Lys

595 600 605

Asp Xaa Gly Xaa Xaa Thr Xaa Asp Ser Val Xaa Leu Ile Pro Ile Pro

610 615 620

Xaa Ser Ala Ala Lys Gly Xaa Gln Xaa Arg Gly Gln Gly Gln Gly Arg

625 630 635 640

Ser Leu Ala Ser Ser Xaa Ser Leu Gln Asp Gln Phe Ala Ala Leu Glu

645 650 655

Lys Glu Leu Gly Xaa Leu Asn Ile Cys Thr Leu Ser Ser Asp Ser Trp

660 665 670

Ser Lys Asp Trp Thr Tyr Thr Val Pro Tyr Xaa Leu Arg Lys Ile Ser

675 680 685

Thr Thr Tyr Ile Gly Thr Val Xaa Lys Ile Asn Xaa Ile Xaa Gly,

690 695 700

wherein

Xaa at position 12 is Met, Leu, Ile or Val;

Xaa at position 34 is Ile or Leu;

Xaa at position 38 is Ile or Leu;

Xaa at position 42 is Glu or Asp;

Xaa at position 43 is Ile or Leu;

Xaa at position 53 is Tyr or Phe;

Xaa at position 55 is Tyr or Phe;

Xaa at position 71 is Gly, Cys or Ala;

Xaa at position 86 is Val or Leu;

Xaa at position 94 is Tyr or Phe;

Xaa at position 97 is Ile or Leu;

Xaa at position 101 is Tyr or Phe;

Xaa at position 103 is Ile, Leu, Gly, Val, Trp, Phe, Thr, Cys, Glu or Arg;

Xaa at position 105 is Met, Gly, Val Leu, Trp, Phe, Pro, Thr, Cys, Asn, Gln or Arg;

Xaa at position 106 is Ile or Leu;

Xaa at position 108 is Gly, Ala, Leu, Ile, Met, Trp, Phe, Ser, Thr, Cys, Tyr, Asn, Asp, Lys or His;

Xaa at position 109 is Ile, Leu, Ala, Val, Leu, Met, Trp, Phe, Pro, Cys, Asn or Glu;

Xaa at position 110 is Glu, Gly, Ala, Val, Leu, Met, Trp, Ser, Thr, Cys, Tyr, Asp or His;

Xaa at position 111 is Tyr, Gly, Ala, Val, Leu, Ile, Met, Trp, Ser, Thr, Cys, Asp, Glu, Lys, Arg or His;

Xaa at position 115 is Asp or Glu;

Xaa at position 119 is Val, Ala, Ile or Leu;

Xaa at position 134 is Ser or Leu;

Xaa at position 137 is Val, Phe, Ala, Leu, Trp, Pro, Ser or Cys;

Xaa at position 139 is Glu or Asp;

Xaa at position 141 is Phe, Leu, Ile, Trp, Ser or Cys;

Xaa at position 144 is Ala, Val, Gly, Ile, Leu or Met;

Xaa at position 148 is Ser, Phe, Thr or Trp;

Xaa at position 152 is Ile, Thr, Leu, Val, Met or Ser;

Xaa at position 155 is Asp or Glu;

Xaa at position 179 is Gly, Val, Trp, Ser, Cys or Arg;

Xaa at position 181 is Ile, Val, Met or Leu;

Xaa at position 182 is Trp, Gly, Ala, Val, Leu, Met, Ser, Cys, Glu or Arg;

Xaa at position 188 is Val or Leu;

Xaa at position 196 is Lys or Glu;

Xaa at position 197 is Thr or Ser;

Xaa at position 201 is Trp, Cys, Tyr or Phe;

Xaa at position 202 is Lys, Asn or Arg;

Xaa at position 203 is Tyr or Phe;

Xaa at position 208 is Glu or Asp;

Xaa at position 214 is Ile or Leu;

Xaa at position 220 is Ile or Leu;

Xaa at position 224 is Tyr or Phe;

Xaa at position 234 is Glu or Asp;

Xaa at position 235 is Val or Leu;

Xaa at position 270 is Ile, Val, Leu or Met;

Xaa at position 296 is Lys or Glu;

Xaa at position 298 is Ala, Glu, Gly or Asp;

Xaa at position 299 is Glu, Gly, Asp or Ala;

Xaa at position 300 is Ile, Val, Ile or Met;

Xaa at position 305 is Asp or Glu;

Xaa at position 317 is Ala, Ser, Gly or Thr;

Xaa at position 323 is Glu or Asp;

Xaa at position 335 is Glu or Asp;

Xaa at position 352 is Glu or Asp;

Xaa at position 359 is Glu, Gly, Ala, Val, Leu, Trp, Phe, Ser, Thr, Lys or Arg;

Xaa at position 360 is Asn, Gly, Val, Leu, Met, Phe, Pro, Thr, Asn, Asp, Lys or His;

Xaa at position 361 is Ser, Gly, Val, Leu or Glu;

Xaa at position 363 is Asp, Gly, Trp or Ser;

Xaa at position 364 is Val, Pro, Ser, Thr, Asn, Glu or Lys;

Xaa at position 365 is Leu, Gly, Ala, Val, Ile, Trp, Phe, Pro, Ser, Thr, Gln, Glu, Arg or His;

Xaa at position 367 is Glu or Lys;

Xaa at position 368 is Gly or Asp;

Xaa at position 370 is Ile, Val, Leu or Met;

Xaa at position 373 is Arg or Ser;

Xaa at position 374 is Asn, Lys, Gln or Arg;

Xaa at position 377 is Leu, Ile, Val or Met;

Xaa at position 384 is Thr, Ala, Ser or Gly;

Xaa at position 385 is Ile, Ser, Leu, Val, Met or Thr;

Xaa at position 388 is Asp or Glu;

Xaa at position 393 is Tyr, Phe or Trp;

Xaa at position 398 is Ala or Val;

Xaa at position 414 is Tyr or Phe;

Xaa at position 418 is Ile or Leu;

Xaa at position 419 is Ser, Asn, Thr or Gln;

Xaa at position 423 is Val or Leu;

Xaa at position 425 is Glu or Val;

Xaa at position 427 is Ile or Val;

Xaa at position 434 is Met or Thr;

Xaa at position 481 is Glu or Asp;

Xaa at position 495 is Asp or Glu;

Xaa at position 509 is Phe, Gly, Ala, Val, Leu, Ile, Met, Trp, Ser, Cys, Tyr, Asn, Asp, Glu or Arg;

Xaa at position 512 is Asn, Ser, Gly, Ala, Leu, Met, Trp, Phe, Ser, Thr, Cys, Gln or Arg;

Xaa at position 514 is Glu, Asp or Arg;

Xaa at position 516 is Gly, Ala, Val, Met, Pro, Thr, Asn, Gln, Asp, Glu or Lys;

Xaa at position 519 is Leu, Gly, Ala, Val, Met, Phe, Pro, Tyr, Gln, Asp or Lys;

Xaa at position 526 is Val or Leu;

Xaa at position 530 is Ile or Leu;

Xaa at position 533 is Val or Ala;

Xaa at position 536 is Ile or Leu;

Xaa at position 538 is Tyr, Phe or Trp;

Xaa at position 543 is Tyr or Phe;

Xaa at position 544 is Lys or Arg;

Xaa at position 547 is Tyr or Phe;

Xaa at position 550 is Tyr, Phe or Trp;

Xaa at position 552 is Asn, Ser, Gln or Thr;

Xaa at position 558 is Phe or Leu;

Xaa at position 600 is Met or Val;

Xaa at position 602 is Met, Ile, Leu or Val;

Xaa at position 607 is Asp or Gly;

Xaa at position 610 is Thr, Lys, Ser or Arg;

Xaa at position 612 is Ile or Thr;

Xaa at position 613 is Leu or Pro;

Xaa at position 615 is Asn or Asp;

Xaa at position 619 is Lys or Arg;

Xaa at position 625 is Tyr, Phe or Trp;

Xaa at position 631 is Ile, Val, Leu or Met;

Xaa at position 633 is Trp or Phe;

Xaa at position 646 is Gln or Arg;

Xaa at position 661 is Asn or Ser;

Xaa at position 683 is Thr, Ala, Ser or Gly; and

Xaa at position 696 is Glu or Asp;

Xaa at position 700 is Ser or Gly; and

Xaa at position 702 is Phe or Ser;

and wherein, 1 to 25 amino acids are optionally deleted from the C-terminus of the polypeptide.

8. The recombinant AfIP-1B polypeptide of claim 5 , wherein the recombinant AfIP-1B polypeptide comprises an amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 76.

9. The recombinant AfIP-1B polypeptide of claim 5 , wherein the recombinant AfIP-1B polypeptide comprises one or more property selected from

a) an amino acid motif as represented by positions 101-105 of SEQ ID NO: 259;

b) an amino acid motif as represented by positions 133-144 of SEQ ID NO: 259;

c) an amino acid motif as represented by positions 177-184 of SEQ ID NO: 259;

d) an amino acid motif as represented by positions 358-365 of SEQ ID NO: 259;

e) an amino acid motif as represented by positions 511-520 of SEQ ID NO: 259;

f) fungicidal activity;

g) insecticidal activity; and

h) a calculated molecular weight of between about 72.5 kD and about 80 kD.

10. A composition, comprising a pesticidally-effective amount of the recombinant AfIP-1B polypeptide of claim 5 .

11. A composition, comprising:

i) an insecticidally-effective amount of

a) a recombinant AfIP-1A polypeptide having at least 85% identity to the amino acid sequence of SEQ ID NO: 2, wherein the AfIP-1A polypeptide comprises the amino acid motifs as represented by positions 15-26 of SEQ ID NO: 257, positions 33-53 of SEQ ID NO: 257, positions 71-84 of SEQ ID NO: 257, and positions 100-107 of SEQ ID NO: 257;

b) a recombinant AfIP-1B polypeptide having at least 95% identity to the amino acid sequence of SEQ ID NO: 4; and

ii) an agriculturally acceptable carrier.

12. The composition of claim 11 , wherein the AfIP-1A polypeptide comprises the amino acid sequence of SEQ ID NO: 257, wherein