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Temperature is an important environmental factor that affects lettuce (Lactuca sativa L.) germination. The present research was conducted to determine the role of seed coverings on lettuce seed germination at high temperature. Five lettuce genotypes were primed in order to bypass thermoinhibitional effects on germination. During germination of primed and nonprimed seeds, imbibition followed a normal triphasic pattern. Primed seeds had higher final water content, a decreased imbibitional phase II, and germinated at 36 °C compared to nonprimed seeds of thermosensitive genotypes, which did not germinate at 36 °C. Puncture tests were conducted to determine the force required to penetrate the whole seed or endosperm of the five genotypes at 24 and 33 °C. `Dark Green Boston', a thermosensitive genotype, had the highest mean resistance (0.207 N) and PI 251245, a thermotolerant genotype, had the lowest (0.139 N). Resistance to penetration of the endopserm of the five genotypes was different at both temperatures. However, three thermotolerant genotypes had lower endosperm resistance than two thermosensitive types. At 36 °C, the penetration force for primed and nonprimed seeds was compared after the first hour of imbibition and 1 hour before radicle protrusion. The force required to penetrate the seed was affected by genotype, seed priming, and duration of imbibition. Puncture force decreased as imbibition time at 36 °C increased in primed and nonprimed seed of each thermotolerant genotype but not in the thermosensitive genotypes. Priming reduced the initial force necessary to penetrate the seed and endosperm in all genotypes. Thus, for radicle protrusion to occur, there must first be a decrease in the resistance of the endosperm layer as evidenced by priming or thermotolerant genotype. Then, the pericarp and integument are sufficiently weakened so that tissue resistance is lower than the turgor pressure of the expanding embryo, allowing germination to be completed.

Lettuce (Lactuca sativa L.) seeds can fail to germinate at temperatures above 24 °C. The degree of thermotolerance is thought to be at least partly related to the environment under which the seed developed. In order to study the effects of temperature during seed development on subsequent germination, various lettuce genotypes were screened for their ability to germinate at temperatures ranging from 20 to 38 °C. Seeds of the selected genotypes `Dark Green Boston' and `Valmaine' (thermosensitive), `Floricos 83', `Everglades', and PI 251245 (thermotolerant) were produced at 20/10, 25/15, 30/20, and 35/25 °C day/night temperature regimes in plant growth chambers. Seeds were germinated on a thermogradient bar from 24 to 36 °C under 12 h light/dark cycles. As germination temperature increased, the number of seeds that failed to germinate increased. Above 27 °C, seeds matured at 20/10 or 25/15 °C exhibited a lower percent germination than seeds that matured at 30/20 or 35/25 °C. Seeds of `Dark Green Boston' and `Everglades' that matured at 30/20 °C exhibited improved thermotolerance over those that matured at lower temperatures. Seeds of `Valmaine' produced at 20/10 °C exhibited 40% germination at 30 °C, but seeds that matured at higher temperatures exhibited over 95% germination. Germination of `Valmaine' at temperatures above 30 °C was not affected by seed maturation temperature. The upper temperature limit for germination of lettuce seed could thus be modified by manipulating the temperature during seed production. The potential thermotolerance of seed thereby increased, wherein thermosensitive genotypes became thermotolerant and thermotolerant genotypes (e.g., PI251245) germinated fully at 36 °C. This information is useful for improving lettuce seed germination during periods of high soil temperature, and can be used to study the biology of thermotolerance in lettuce.

Two random amplified polymorphic DNA (RAPD) markers linked to En, the gene conferring resistance to pea enation mosaic virus in pea, were identified and the DNA fragments were cloned and partially sequenced. Allele-specific associated primers for each cloned DNA fragment were developed and used in screening F₂ populations. One marker, P256₉₀₀, mapped very near Adh-1, about 6 cM from En. The other marker, B500₄₀₀, was located about 8 cM from En on the same side as P256₉₀₀.

Petunia ×hybrida (Hook) Vilm. cv. Mitchell was transformed with an E. coli gene encoding mannitol-1-phosphate dehydrogenase (mtlD). Four plant lines that grew on kanamycin and contained the mtlD transgene were identified. Two of these lines contained high levels of mannitol [high-mannitol lines M3 and M8; mean mannitol = 3.39 μmol·g^-1 dry weight (DW)] compared to nontransformed wild-type plants (0.86 μmol·g^-1 DW), while two lines had mannitol levels similar to wild-type plants (low-mannitol lines M2 and M9; mean mannitol = 1.05 μmol·g^-1 DW). Transgenic and control plants were subjected to chilling stress (3 ± 0.5 °C day/0 ± 0.5 °C night, 12-hour photoperiod and 75% relative humidity) to evaluate the role of mannitol in chilling tolerance. Based upon foliage symptoms and membrane leakage after a 3-week chilling treatment, the high-mannitol containing lines, M3 and M8, were more tolerant of chilling stress than the low-mannitol containing transgenic lines, M2 and M9, and wild-type. Under nonchilling conditions mannitol was the only carbohydrate that differed among transgenic lines, but all carbohydrates were present. When subjected to chilling stress, mannitol levels dropped by 75%, sucrose by 52%, and inositol by 54% in the low-mannitol lines (M2 and M9). In M3 and M8, the high-mannitol lines, mannitol levels decreased by 36%, sucrose by 25%, and inositol by 56%, respectively. Raffinose increased 2- to 3-fold in all lines following exposure to low-temperature chilling stress. In the higher mannitol lines only 0.04% to 0.06% of the total osmotic potential generated from all solutes could be attributed to mannitol, thus its action is more like that of an osmoprotectant rather than an osmoregulator. This study demonstrates that metabolic engineering of osmoprotectant synthesis pathways can be used to improve stress tolerance in horticultural crops.