NE124: Genetic Manipulation of Sweet Corn Quality and Stress Resistance
- Duration:
- October 01, 1999 to September 30, 2005
- Administrative Advisor(s):
-
Max J. Pfeffer
(NY.C)
- NIFA Reps:
-
AnnMarie Thro
Statement of Issue(s) and Justification:
Sweet corn represents one of the important vegetable crops in the United States. Annual sweet corn production in this country occurs on 800,000 acres (appx.). Additionally, it is an important seed export for the U.S. inasmuch as the U.S. supplies most of the seed for production for the rest of the world. Unfortunately, much of the enormous investment in field corn research is not directly relevant to the contemporary production problems of sweet corn. Among these sweet-corn-unique problems are the need to: reinforce the presently weak germplasm-base representing sweet corn types; reconcile, inter alia, the vulnerability of sweet-type seed to soil borne and leaf pathogens; and resolve the genetics of sweet corn quality as seed and as a food.
Related, Current, and Previous Work:
A. Germplasm Resources for Sweet Corn Improvement.Of the hundreds of open-pollinated varieties existing in 1900, today only about 50 remain. Most of the remaining varieties have been evaluated for germination and seedling growth under cold temperatures" and reaction to a number of pathogens". Hotchkiss et al." found substantial variation for the ability to grow under cold temperatures among 35 varieties. A few of the varieties grew very well equaling the performance of Mexican high altitude sweet corn and exceeding Corn Belt Dent germplasm that had been selected for cold tolerance". Pataky et al.14 screened 36 varieties for reaction to Stewart's wilt (Erwinia stewartii), common rust (Puccinia sorghi), northern leaf blight (NLB) (Exserohilum turcicum) , and southern leaf blight (SLB) (Bipolaris maydis). Modern commercial germplasm was more resistant to all four diseases than were the open-pollinated varieties84 A few of the open-pollinated varieties were resistant to certain diseases and may represent novel sources of resistance alleles84.
Many sweet corn breeders have resisted using non-sweet germplasm, warning against the difficulties in retaining table quality factors and specific raw product characteristics important in sweet corn61,72,101 However, high levels of resistance to many pests is available in non-sweet corn germplasm. Genes for resistance to common rust and maize dwarf mosaic virus from Corn Belt Dent germplasm have been incorporated into sweet corn",3'.'0. Sweet corn inbreds with partial resistance to NLB have been developed using tropical corn varieties". Davis et al." developed a rust resistant population using 11 sugaryl (sul) inbreds and 17 Latin American varieties. The Corn Belt Dent inbred B52 was used as a source of genes for resistance to European corn borer (ECB) (Ostrinia nubilalis)37,69 and Latin American germplasm has also been examined as sources of ECB resistance". In developing sweet corn for the tropics, Brewbaker has emphasized tropical germplasm sources, because most North American sweet corn germplasm lacks the pest resistance and stress tolerance needed to survive under tropical conditions with minimal inputs 15,16,17,18.
Common rust causes serious yield and quality reductions throughout much of the sweet corn growing area. Two forms of resistance are available; general or partial, in which the size and number of pustules are reduced, and specific, in which a hypersensitive reaction restricts pustule development'03. Hybrids with high levels of partial resistance are available"0g'09'. Specific or single gene resistance conditioned by the RpI locus is available in many hybrids. The RpI locus is a complex locus consisting of duplicated sequences62, 65, 64, 111 ". A high frequency of unequal crossing over has generated a large number of alleles64, 92 and new allele combinations have been fixed and released". The allele most frequently used today is Rpl -d. However, other alleles at Rpl as well as the c allele at Rp3 have some utility''088. Resistance due to alleles at Rpl and Rp3 is usually dominant. A case of overdominance has been reported at another rust resistance locus, Northern leaf blight can cause economic damage in sweet corn throughout the eastern half of the USA. Both partial and single gene forms of resistance exist for NLB". However, races of the pathogen are able to overcome all commonly used single genes '9''02,103_ The polygenic form of resistance is effective regardless of races present'03.
Kernel and ear rots caused by Fusarium, Diplodia, and Gibberella are a problem in sweet corn seed production. Fusarium kernel rot is often associated with insect damage and differences in susceptibility to Fusarium exist among sweet corn inbreds55 In supersweets, reduced germination and seedling dieback can result from Fusarium infection of the developing kernels". Maternal effects are important in resistance to Fusarium, suggesting that the site of resistance may be the pericarp or the silk". Under high humidity ear and kernel rots are intensified. Gibberella zeae ear rot has appeared on sweet corn grown for processing"," 9. Under certain conditions G. zeae can produce significant levels of toxins prior to fresh ear harvest.
A number of synthetic and composite populations have been developed by university and USDA researchers. In developing composite `AS 1 R', Rubino and Davis" performed 10 cycles of mass selection for earliness and pest resistance. Three populations have undergone three cycles of full-sib recurrent selection for resistance to common rust with decreases in rust damage per cycle of 8, 12, and 13%'. MINN I 1 responded to a divergent selection program for endosperm phenotype'023. Brewbaker has developed populations for direct use as open-pollinated varieties15, 16, 17,18 Brewbaker has used shrunken2 (sh2), brittle] (btl ), and brittle2 (bt2) in different varieties, all of which are adapted to the tropics.
The use of molecular markers for marker assisted selection programs has been suggested by many workers42,106. In sweet corn, RFLPs have been used to classify and evaluate the variation among publicly available sweet corn inbreds46, and to identify the chromosomal location of some quantitative trait loci (QTL)6,7,47. RFLPs have been used to map single genes such as sel113 Preliminary data from Juvik's group at Illinois indicates that MAS can be effective in improving germination.
B. Biochemistry and Physiology.
Sweet corn flavor is determined in part by the amount of sugar in the endosperm. Starch synthesis in the corn endosperm has been the subject of much research. Reviews (many by NE124 members) on starch biosynthesis and the genetic modification of endosperm carbohydrates have been published' Most mutants used in sweet corn improvement increase sugar content and decrease starch content. One mutant, sul, elevates the level of water soluble polysaccharide (WSP) (phytoglycogen) in addition to increasing sugar content".3o. Eight starch synthesis mutants have been used commercially. For all but sul the basic science and initial development and commercialization was done by NE-124 cooperators. Most of these genes have been cloned and sequenced and their specific enzymatic lesions are known 8'11,22,44,48,50,67,90,98,99,100,112. Starch synthesis mutants are divided into two classes based on their effects on endosperm composition". The class 1 mutants accumulate sugars at the expense of starch and have greatly decreased total carbohydrates in the mature seed 130g2. At 18 to 21 days after pollination these mutants have 4 to 8 times the total sugar found in nonmutant corn20,30,26,58,59,68,75,82,116. Due to high sugar levels, class 1 mutants are used independently in sweet corn varieties. For processing, sh2 is currently the second most widely used endosperm type after sul, while for many fresh uses, usage of sh2 has surpassed that of sul.
Class 2 mutants alter the types and amounts of polysaccharides produced '3, The alleles ae], dul, and wxl generally result in slightly less starch in the mature kernel than nonmutant types '3,30,31,82. These 3 mutants result in small increases in sugar content and do not make acceptable sweet corn when used singly. However, complementary gene action of certain triple combinations of class 2 mutants results in sugar levels equal to those found in class I MUtants29,31,57. Commercial hybrids having the triple recessive genotype ae] dul wxl have been released 45. The sul allele results in greatly increased levels of WSP82,31. WSP is a highly branched polysaccharide 12 and gives sul endosperm the smooth texture and creaminess, characteristic of traditional sweet corn varieties13,31,33,11. While high levels of WSP appear to be unique to sul, elevated levels are maintained when sul is combined with either wxl, dul, or btl3,19,29,52, The sugary enhancerl (sel) allele"3 when in combination with homozygous sul, results in sugar levels near those of sh2 and WSP levels similar to unmodified SU 15'43,a9. This results in a high quality, sweet, creamy endosperm. It appears that a number of recessive modifiers are required to attain high quality sul sel hybrids"4.
Many important endosperm genes have extensive allelic series13,27,52. Phenotypic expression of these alleles range from smooth seeded, normal appearing kernels to ones so defective as to be lethals. The presence of some normal appearing alleles can only be detected when another gene is present27 It is not known if all the hybrids of a given endosperm type have the same allele at that locus. However, it has been suggested that certain alleles of sh2 and bt2 may have better seed quality, while maintaining adequate levels of sugar for acceptable flavor12,11
Many endosperm mutants have poor germination and seedling vilgor28,80,89,93. Combinations of endosperm mutants usually result in even lower germination and seed vigor93,95. Mutants that accumulate high sugar levels have reduced germination relative to normal corn and other mutants, especially in cold Soils 110,118,122 Hybrids containing sel generally have reduced germination relative to SU140. Understanding the causes of poor field emergence in high-sugar types and developing techniques to improve it are crucial in making high-sugar varieties commercially acceptable.
Genetic background strongly affects emergence, and background by endosperm mutant interactions are also important in determining emergence and seedling vigor 4'40,93,95,110,118,122. Significant effects of genetic background on germination indicate improvement of germination is possible through selection. A sh2 population mass selected for improved emergence and seed weight had increased cold and warm9 Similar changes have been observed in a sul ponulation24. Significant background by endosperm type interactions indicate that breeders can not assume that backcrossing a new endosperm mutant into a line with good seed quality will result in a line that germinates well".
Reduced emergence is affected by both genetic and environmental conditions, both during seed production and at planting","'. Seed weight of sh2 is 33 to 50% that of sul 96'"g and seed weight is correlated to percent germination 40". Reduction in seed weight is a function of greatly reduced starch levels relative to other types of corn105, 107 Thus, starch levels are related to germination and seed vigor both among endosperm types'o' and within sh2". On a percent of dry weight basis, sh2 seed has high levels of sugars 104,101. In an extensive study on effects of recurrent selection for improved germination in a sh2 population, sugar levels were not directly related to germination". However, due to the osmotic potential caused by high sugar levels, sh2 corn dries very slowly and maturing seed is thus more susceptible to frost damage during maturation114. Slow drydown rates also may be responsible for the higher incidence of seed-borne Fusarium moniliforme Sheldon in sh2 seed"g. Infection by F. moniliforme and other pathogens can greatly reduce germination and seed vigor". While slow drydown rates of sh2 seed may be responsible for increased infection of seed by F. moniliforme, increased infection is not directly related to high sugar concentration16.
Low carbohydrate concentration of sh2 results in severe shrinking of the endosperm as it dries, which creates a number of structural problems for the seed""', including cracking of the pericarp. The pericarp is a barrier to pathogens and water movement. Damaged pericarp greatly reduces germination in all endosperm types".
Starch granule degradation in the subaleurone endosperm was significantly. less in a sh2 variety compared with non-isogenic sul, sul sel, and normal varieties". Thus, the availability of food for the germinating seed may be reduced. Both respiration rate and adenosine-triphosphate (ATP) levels have been studied and neither can account for the poor seed vigor in sh2 corn"o0"g.
Effects of soil-borne pathogens on germination of sh2 seed and the need for chemical seed treatments were recognized early in the development of sh2 as a crop'002'. Reliable fungicides are required for high-sugar corns to be an viable crop. Fungicide combinations have been observed to increase germination under cold test performance three to four fold relative to the untreated check'''. Three to five different fungicides plus an insecticide are commonly applied to commercially treated sweet corn seed"0"O.
C. Insects, Diseases, and Weeds.
As a result of concerns related to human health, nontarget organisms, and the environment, the reduction of pesticide use in US agriculture is an important goal. A means to achieve this goal is through the development and adoption of the integrated pest management (IPM) strategy, an ecologically based strategy that promotes the use of non-chemical control tactics. Pesticides are a key component of IPM, but they are only used when all other options fail to control the pest. The continued availability of pesticide options is in jeopardy due to the development of resistant pests and changes in pesticide regulations. The recent enactment of the Food Quality Protection Act may result in the loss of many classes of certain pesticides. Alternatives be developed for sweet corn.
The ECB is the most serious insect pest in the major processing states, while the corn earworm (Heliothis zea) and fall armyworm (Spodoptera frugiperda) cause significant damage in other regions. Inheritance of resistance to these pests has been the subject of intense investigation in field corn39, and breeding for ECB and corn earworm resistance is a major effort in public sweet corn programs"","'. However, the genetic variability for resistance within elite sweet corns is low". The development of sweet corn possessing the Bacillus thuringiensis (Bt) toxin has the potential to revolutionize control of lepidopteran pests of sweet corn"0'6. Key to the long-term effectiveness of Bt sweet corn however, is the development and adoption of a resistance management program. NE-124 cooperators are well positioned to address this need". Other insect pests of sweet corn include the corn leaf aphid (Rhopalosiphum maidis), corn flea beetle (Chaetocnema pulicaria, the vector of Stewart's wilt), and sap beetles (Nitidulidae). With the availability of genetically engineered Bt sweet corn, infestations of pests previously controlled by insecticides applied for control of ECB may become more common. Herbicides constitute 60 to 70% of the pesticide use in U.S. agriculture. With enactment of the Food Quality Protection Act the availability of a wide spectrum of herbicides may be in jeopardy. Efforts need to be made to evaluate and optimize existing and new herbicides and find cost-effective alternatives. Numerous strategies for herbicide reduction are being researched across the country. Of particular importance to sweet corn, weed management tactics such as cultivation, reduced tillage, banded applications, and cover crops need to be developed. While published literature includes a significant amount of efficacy and phytotoxicity information on the individual herbicides in field corn, there is little published on sweet corn and nothing that relates to planning a total herbicide program. Sweet corn is planted at different times than field corn, emerges slowly and at the same time as many of the summer annual weeds, and is therefore far less competitive than field corn. Need for effective weed control is more critical in sweet corn than in field corn if high yields are to be maintained. An additional problem with sweet corn is the number of varieties, both for fresh market and processing. There is documented evidence of increased herbicide injury due to the extreme differences in vigor between the traditional, sugarenhanced, and supersweet varieties. Research needs to be conducted to determine a screening strategy for herbicide tolerance particularly in the sugar-enhanced and the supersweet varieties and to develop economical weed management programs that are not based on atrazine, or which minimize its use.
Objectives
- Acquire, characterize, and preserve existing sweet corn germplasm, and use traditional and novel techniques to create additional genetic variability.
- Determine the genetic, biological, and physiological mechanisms in sweet corn regulating seed and food quality.
- Reduce environmental impacts of sweet corn production, with a focus on host plant resistance and abiotic stress tolerance.
