Nematospora seed rot and lint stain: Distribution, Importance and Association with Hemiptera Alois Bell, Juan Lopez, Jr., Enrique Medrano, Jack Bacheler,

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Nematospora seed rot and lint stain: Distribution, Importance and Association with Hemiptera Alois Bell, Juan Lopez, Jr., Enrique Medrano, Jack Bacheler, Jeremy Green, Phillip Roberts, Robert Kemerait, Jr., James Marois, David Wright, and Robert Nichols USDA-ARS, College Station, TX University of North Carolina, Raleigh, NC Clemson University, Clemson, SC University of Georgia, Tifton, GA University of Florida, Quincey, FL Cotton Incorporated, Cary, NC

Determine the possible importance and etiology of infectious yeast in seed rot, internal boll rot, and tight lock of cotton. Objective

1. Belong to four closely related genera and readily produce 2-celled, needle-shaped ascospores. 2. Ashbya and Eremothecium are filamentous, whereas Holleya and Nematospora are dimorphic. Recent studies place all in Eremothecium. 3. Specifically associated with heteropterous insects, and are restricted to warmer climates. 4. Cause diseases of many families of angiosperms, usually attacking fruits or seeds pierced by bugs during feeding. Important crop hosts include cotton, okra, soybean, green bean, cowpea, citrus, pistachio, tomato, and mustard. Introduction to Plant Pathogenic Yeast

Nematospora coryli on TSA medium

Nematospora coryli on PDA medium

Close-up of Nematospora coryli colonies

Colonies showing hyphae Close-up of Nematospora coryli colonies

Nematospora coryli: A – vegetative budding cells, B – hyphae with ascus, C – asci, D – ascospores

Bud cells, hyphae, and ascospores of Nematospora coryli

Symptoms 1. Prior to boll opening: lint is first stained yellow and later reddish-brown and may become dry. Seed coats are discolored and embryos are killed or their development is severly inhibited. 2. At boll opening: sutures may not split completely, locks do not fluff normally and infections by secondary fungi are common, sometimes blackening the lock. 3. After opening: cotton locks are stained tan to reddish brown. Seed is shriveled with discoloration of fuzz and absence of black pigment in seed coat.

Florida lock infected with Nematospora coryli Lock cross-section

Infected lock showing staining and restricted opening

Comparison of locks infected with bacteria or yeast

Secondary Alternaria and Cladosporium infections of locks affected by N. coryli

Seed rot and shriveling caused by N. coryli InfectedControl

Occurence of Nematospora in Field and Greenhouse Bolls* Punctured by Stink Bugs in 2005 Source of Bolls No. Bolls Observed No. With Punctures No. With Nematospora No. With >10 6 Bacteria Field Bolls Greenhouse Bolls* Greenhouse Bolls* (SGSB collected from millet on 9/15 & 10/6) Females Males Nymphs12220 Greenhouse Bolls* Greenhouse Bolls* (SGSB collected from light trap on 10/5) Females10313 Males6303 * Single insects caged over bolls for 8 days. Microbial content measured 12 days after insects removed.

Frequency of Caged Feral Insects Transmitting Nematospora and Bacterial Pathogens to Bolls in 2006 No. of BollsNo. With Pathogen Insect*Source † TotalPuncturedNematosporaBacteriaBoth SGSBMillet BSBMillet BSBP Trap LFPBMillet98743 Total * SGSB = Southern green stink bug; BSB = Brown stink bug; LFPB = Leaf-footed plant bug. † Collected from pearl millet by hand or from pheromone traps.

Effect of Insecticides on Insect Punctures and Boll Infections by Nematospora and Bacteria in 2006 % Bolls* Punctured % Punctured Bolls With StateTreatment NematosporaBacteriaBoth NCNone60 (6.0) Insecticide6 (1.0)060 SCNone26 (2.0)18270 Insecticide4 (1.0)5000 GANone94 (25.1) Insecticide26 (10.5) FLNone62 (21.9)62238 Insecticide60 (10.6)55246 * Based on 50 bolls; ( ) indicates mean number of punctures.

Effects of Nematospora coryli (Seed and Fiber Development in Lock)

Effects of Nematospora coryli on Seed

Effects of Nematospora coryli (Array of Cotton Seed Weights - ST 474) * Mean of >400 bolls from 98 cultivars.

Effects of Nematospora coryli (Array of Cotton Seed Weights - DP 393) * Mean of >400 bolls from 98 cultivars.

Effect of Boll Age on Damage Caused by Nematospora Boll Age at Inoculation* (Days) % Decrease in Weight LockFiberSeed * Fifty bolls from four cultivars.

Effect of Boll Age on Damage Caused by Nematospora (...continued) Boll Age at Inoculation* (Days) % Decrease in Weight LockFiberSeed * Fifty bolls from four cultivars.

Effect of Boll Age on Infection and Colonization of Nematospora coryli Transmitted by Feeding of Southern Green Stink Bugs Infested Boll Age (Days) Number of Bolls* TotalInfected >10,000 cfu/gm Total * Bolls examined 12 days after single feral insects from pearl millet were caged over bolls of specified ages.

Cultivar Reaction to Nematospora coryli

Cultivar Reaction to Nematospora % Reduction in Fiber Weight Susceptible CultivarsResistant Cultivars ST 5242 BR (80.1)Phyt 710 R Acala (13.7) ST 5599 BR (69.3)DP 110 RF (14.5) NG 3273 B 2 RF (63.8)BW 6896 B 2 F (15.9) BW 2038 B 2 F (63.6)FM 966 LL (16.9) BW 3255 B 2 F (61.6)DP 444 BG/RR (17.6) NG 3550 RF (60.3)AFD 5065 B 2 F (18.9) FM 960 RR (59.8)CPCSD Acala Fiesta (20.6) DP 164 B 2 RF (59.0)ST 6622 RF (21.4) Phyt 370 WR (58.2)BW 8245 B 2 F (24.8) DP 167 RF (58.1)AFD 5062 LL (24.9) ST 4357 B 2 RF (54.8)FM 958 LL (25.9) ST 4700 B 2 RF (54.8)CG 3020 B 2 RF (26.6)

Conclusions 1. Nematospora occurs in many bolls punctured by bugs during feeding. 2. A high percentage of feral insects carry Nematospora which may persist and be transmitted repeatedly. 3. Most cultivars are highly susceptible to Nematospora and suffer loses of 40-60% of the fiber, as well as reducing seed quality. 4. Insect control is the best way to prevent infection, although improved cultivar resistance may be possible.