Development of novel highly chemoprotectant crucifer germplasm

Abstract

A method for producing crucifer germplasm, in particular broccoli and cauliflower germplasm, with high levels of cancer chemoprotectant agents which comprises screening a population of plants for glucosinolate profiles and selecting an individual plant with an enhanced alkyl/indole glucosinolate ratio. Subsequent breeding with a selected plant results in varieties, lines, and plants with desirable glucosinolate profiles. A method of growing crucifer sprouts to enhance chemoprotectant activity of the sprouts by germinating the seeds in a limited water volume. A method for producing crucifer germplasm which produces seeds with reduced leakage of seed material which comprises screening a population of plants for production of seeds with reduced leakage and selecting an individual plant which produces such seeds.

We claim:  
     
         1 . A method for selecting a plant comprising: 
 (a) measuring the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of a plant, and    (b) selecting the plant if the ratio is a specific number.    
     
     
         2 . The method of  claim 1 , wherein the plant is a double haploid plant.  
     
     
         3 . The method of  claim 2 , wherein the double haploid plant is obtained by an anther culture technique.  
     
     
         4 . The method of  claim 1 , wherein the plant is a mutagenized plant.  
     
     
         5 . The method of  claim 4 , wherein the plant was mutagenized with a chemical mutagen before (a).  
     
     
         6 . The method of  claim 5 , wherein the chemical mutagen is ethylmethane sulfonate.  
     
     
         7 . The method of  claim 1 , wherein the plant is a crucifer.  
     
     
         8 . The method of  claim 7 , wherein the plant is a broccoli plant.  
     
     
         9 . The method of  claim 8 , wherein the plant belongs to a cultivar selected from the group consisting of Saga, DeCicco, Everest, Emerald City, Packman, Corvet, Dandy Early, Emperor, Mariner, Green Comet, Green Valiant, Arcadia, Calabrese Caravel, Chancellor, Citation, Cruiser, Early Purple Sprouting Red Arrow, Eureka, Excelsior, Galleon, Ginga, Goliath, Green Duke, Greenbelt, Italian Sprouting, HiSierra, Late Purple Sprouting, Late Winter Sprouting White Star, Legend, Leprechaun, Marathon, Mariner, Minaret (Romanesco), Paragon, Patriot, Premium Crop, Rapine (Spring Raab), Rosalind, Salade (Fall Raab), Samurai, Shogun, Sprinter, Sultan, Taiko, and Trixie, and Viking.  
     
     
         10 . The method of  claim 8 , wherein the plant belongs to a cultivar selected from the group consisting of Viking, Everest, HiSierra, and Marathon.  
     
     
         11 . The method of  claim 7 , wherein the plant is a cauliflower plant.  
     
     
         12 . The method of  claim 11 , wherein the plant belongs to a cultivar selected from the group consisting of Alverda, Amazing, Andes, Burgundy Queen, Candid Charm, Cashmere, Christmas White, Dominant, Elby, Extra Early Snowball, Fremont, Incline, Milkyway Minuteman, Rushmore, S-207, Serrano, Sierra Nevada, Siria, Snow Crown, Snow Flake, Snow Grace, Snowbred, Solide, Taipan, Violet Queen, White Baron, White Bishop, White-Contessa, White Corona, White Dove, White Flash, White Fox, White Knight, White Light, White Queen, White Rock, White Sails, White Summer, White Top, and Yukon.  
     
     
         13 . A method for producing a progeny plant comprising: 
 (a) measuring the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of a potential parent plant,    (b) selecting the potential parent plant if the ratio is a specific number, and    (c) producing a progeny plant from the selected plant.    
     
     
         14 . The method of  claim 13 , 
 wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the progeny plant is higher than the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the selected plant, and    wherein the tissue of the progeny plant and the tissue of the selected plant is of the same type.    
     
     
         15 . A progeny plant produced by the method of  claim 13 , 
 wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the progeny plant is higher than the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the selected plant, and    wherein the tissue of the progeny plant and the tissue of the selected plant is of the same type.    
     
     
         16 . The progeny plant of  claim 15 , wherein the ratio in the tissue of the progeny plant is at least 10% greater than the ratio in the tissue of the selected plant.  
     
     
         17 . The progeny plant of  claim 16 , wherein the ratio in the tissue of the progeny plant is at least 20% greater than the ratio in the tissue of the selected plant.  
     
     
         18 . The progeny plant of  claim 17 , wherein the ratio in the tissue of the progeny plant is at least 30% greater than the ratio in the tissue of the selected plant.  
     
     
         19 . The progeny plant of  claim 18 , wherein the ratio in the tissue of the progeny plant is at least 50% greater than the ratio in the tissue of the selected plant.  
     
     
         20 . The progeny plant of  claim 19 , wherein the ratio in the tissue of the progeny plant is at least 100% greater than the ratio in the tissue of the selected plant.  
     
     
         21 . The progeny plant of  claim 20 , wherein the ratio in the tissue of the progeny plant is at least 10 times as great as the ratio in the tissue of the selected plant.  
     
     
         22 . The progeny plant of  claim 21 , wherein the ratio in the tissue of the progeny plant is at least 100 times as great as the ratio in the tissue of the selected plant.  
     
     
         23 . The progeny plant of  claim 16 , wherein the ratio in the tissue of the progeny plant is between 10% greater than and 100% greater than the ratio in the tissue of the selected plant.  
     
     
         24 . The progeny plant of  claim 23 , wherein the ratio in the tissue of the progeny plant is between 20% greater than and 80% greater than the ratio in the tissue of the selected plant.  
     
     
         25 . The progeny plant of  claim 24 , wherein the ratio in the tissue of the progeny plant is between 30% greater than and 60% greater than the ratio in the tissue of the selected plant.  
     
     
         26 . The progeny plant of  claim 21 , wherein the ratio in the tissue of the progeny plant is between 10 times as great and 100 times as great as the ratio in the tissue of the selected plant.  
     
     
         27 . The progeny plant of  claim 26 , wherein the ratio in the tissue of the progeny plant is between 25 times as great and 75 times as great as the ratio in the tissue of the selected plant.  
     
     
         28 . The progeny plant of  claim 15 , wherein the progeny plant is produced by selfing the selected plant.  
     
     
         29 . The progeny plant of  claim 15 , wherein the progeny plant is a hybrid plant.  
     
     
         30 . A cruciferous plant, wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is greater than 6.  
     
     
         31 . The plant of  claim 30 , wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is greater than 10.  
     
     
         32 . The plant of  claim 31 , wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is greater than 25.  
     
     
         33 . The plant of  claim 32 , wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is greater than 50.  
     
     
         34 . The plant of  claim 33 , wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is greater than 100.  
     
     
         35 . The plant of  claim 34 , wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is greater than 1000.  
     
     
         36 . The plant of  claim 30 , wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is between 6 and 1000.  
     
     
         37 . The plant of  claim 36 , wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is between 10 and 500.  
     
     
         38 . The plant of  claim 37 , wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is between 25 and 250.  
     
     
         39 . The plant of  claim 38 , wherein the ratio of alkyl glucosinolate to indole glucosinolate in a tissue of the plant is between 50 and 100.  
     
     
         40 . The plant of  claim 30 , wherein the plant is a broccoli plant.  
     
     
         41 . The plant of  claim 30 , wherein the plant is a cauliflower plant.  
     
     
         42 . A method for selecting a seed or sprout comprising: 
 (a) adding a quantity of water to a seed or sprout,    (b) measuring the concentration of a glucosinolate in the leachate of the seed or sprout, and    (c) selecting the seed or sprout if the concentration is a specific value.    
     
     
         43 . A method for producing a progeny plant comprising: 
 (a) adding a quantity of water to a seed or sprout,    (b) measuring the concentration of a glucosinolate in the leachate of the seed or sprout,    (c) selecting the seed or sprout if the concentration is a specific value, and    (d) producing a progeny plant from the selected seed or sprout.    
     
     
         44 . A method for inhibiting the growth of a microbe in an environment capable of sustaining the microbial growth comprising administering to the environment a microbial growth-inhibiting amount of a cruciferous seed leachate or a cruciferous sprout leachate.  
     
     
         45 . The method of  claim 44 , comprising administering to the environment a microbial growth-inhibiting amount of a cruciferous sprout leachate.  
     
     
         46 . The method of  claim 44 , wherein the method prevents the microbial growth.  
     
     
         47 . The method of  claim 44 , wherein the microbe is a bacterium.  
     
     
         48 . The method of  claim 44 , wherein the microbe is Escherichia spp.  
     
     
         49 . The method of  claim 44 , wherein the microbe is  Escherichia coli.    
     
     
         50 . The method of  claim 44 , wherein the microbe is Staphylococcus spp.  
     
     
         51 . The method of  claim 44 , wherein the microbe is  Staphylococcus aureus.    
     
     
         52 . The method of  claim 44 , wherein leachate is a broccoli leachate.  
     
     
         53 . The method of  claim 44 , wherein the leachate is a cauliflower leachate.  
     
     
         54 . The method of  claim 44 , wherein the leachate contains a glucosinolate.  
     
     
         55 . A method for preparing sprouts comprising: 
 (a) providing a quantity of seeds,    (b) adding to the seeds between 0.6 and 1.0 g of water per g of seeds,    (c) allowing the watered seeds to germinate and form sprouts, and    (d) adding to the sprouts between 5 and 9 mg of water per sprout per 24 h for 24 to 120 h, thereby preparing the sprouts.    
     
     
         56 . The method of  claim 55 , wherein (b) comprises adding to the seeds between 0.7 and 0.9 g of water per g of seeds.  
     
     
         57 . The method of  claim 55 , wherein (b) comprises adding to the seeds 0.8 g of water per g of seeds.  
     
     
         58 . The method of  claim 55 , wherein (d) comprises adding to the sprouts between 6 and 8 mg of water per sprout per 24 h for 24 to 120 h.  
     
     
         59 . The method of  claim 55 , wherein (d) comprises adding to the sprouts 7 mg of water per sprout per 24 h for 24 to 120 h.  
     
     
         60 . The method of  claim 55 , wherein (d) comprises adding to the sprouts between 5 and 9 mg of water per sprout per 24 h for 48 to 96 h.  
     
     
         61 . The method of  claim 55 , wherein (d) comprises adding to the sprouts between 5 and 9 mg of water per sprout per 24 h for 72 h.  
     
     
         62 . The method of  claim 55 , wherein the sprouts are cruciferous.  
     
     
         63 . The method of  claim 55 , wherein the sprouts are broccoli sprouts.  
     
     
         64 . The method of  claim 55 , wherein the sprouts are cauliflower sprouts.