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Kenneth W. Mudge and Chin-Chang Chu

In vitro asymbiotic seed germination, subculture, and outplanting of orchids is presented as a laboratory exercise suitable for students of plant propagation or tissue culture. Dendrobium antennatum (Lindley), Phalaenopsis (Blume) white hybrid, or both, are used in this exercise because they flower predictably in the greenhouse, are reliable for seed production, and germinate and grow rapidly in vitro. The exercises can be used to instruct students in the skills involved in orchid seed sterilization, sowing, and culture, as well as instruct students in the unique features of orchid reproductive biology and symbiosis. A schedule is suggested for stock plant flower pollination, capsule harvest, seed sowing, and seedling subculture so that the necessary plant material is available for students to sow, subculture, and outplant seedlings during a single laboratory session.

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K.S. Mayberry, J.A. Principe, and W.L. Schrader

The artichoke, Cynara scolymus, is normally propagated by cuttings from mother plants; however, it is possible to produce some types of artichokes from seed. Methods used for producing open-pollinated seed of onion and carrots may be suitable for producing artichoke seed. Outcrossing in artichokes occurs because of differences in maturity of the staminate and pistillate phases within flowers. Producing artichoke seed by simple inbreeding techniques is usually not successful because of vigor loss and low pollen production, low seed production, and late maturity of progeny. Outcrossing is the preferred method of creating a new variety. The cultivar `Imperial Star' was developed by crossing a thornless French line with an Italian line that had sharp woody spines, and a uniform, olive-green color. The French line was a bright green with some light purple at the base of the bracks. The F1 generation from this cross had good hybrid vigor, and produced abundant seed and pollen. The F2 generation segregated widely with many recombinant types that neither parent showed (e.g., extreme thorniness of leaves and petioles). Two plants were selected for sibling pollination. Subsequent generations of siblings within this type produced higher percentages of the desired type—glossiness, earliness, and high seed yield. Subsequent sibling crossing led to the selection of `Imperial Star', PVP. 9000179.

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R.A. Stern, S. Gazit, R. El-Batsri, and C. Degani

Fruits produced in two orchards, each consisting of adjacent blocks of `Floridian' and `Mauritius' lychee (Litchi chinensis Sonn.), were unequivocally identified as selfed or outcrossed by phosphoglucose isomerase (PGP; EC isozyme analysis. The average rate of hybrid production in each orchard was 69% and 87% for `Floridian' and 17% and 65% for `Mauritius', respectively. The percentage of hybrids produced on trees adjacent to those of the other cultivar was invariably significantly higher than that produced on the more distant trees. However a significant correlation between hybrid percentage and proximity to the other cultivar, as well as between hybrid percentage and yield, was found only for `Floridian' in one of the orchards. A significant correlation was found between pollen source and the weights of fruits and seeds in both cultivars. Fruits originating from cross-pollination were heavier and contained heavier seeds than selfed fruits. The most pronounced effect of pollen parent on seed weight was found in `Floridian', which appears to exhibit inbreeding depression.

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Troy A. Larsen* and Christopher S. Cramer

New Mexico onion production will begin using mechanical harvesters in the near future in order to stay competitive in today's market. Past onion breeding objectives have focused on improving onions for hand harvesting instead of mechanical harvesting. Our breeding program is starting to evaluate germplasm for bulb firmness. The objectives of this study were to evaluate hybrid lines for their bulb firmness, to compare two methods of measuring bulb firmness, and to compare bulb firmness using two different production schemes. Bulb firmness of spring-transplanted and spring-seeded intermediate-day hybrid breeding lines was measured using a digital FFF-series durometer and a subjective rating of firmness achieved by squeezing bulbs. Bulbs were rated on a scale of 1 (soft) to 9 (hard). In general, these hybrid lines produced very firm to hard onions whether the lines were transplanted or direct-seeded. Bulb firmness of these lines measured with the durometer was greater when the lines were direct-seeded (74.9) than when transplanted (73.5). Conversely, when firmness was measured with our subjective rating, transplanted onions exhibited slightly greater firmness (8.9) than direct-seeded onions (8.8). For both transplanted and direct-seeded onions, durometer readings were weakly correlated in a positive fashion with our subjective rating. In general, durometer readings gave a greater spread in firmness measurements with a range of 69.6 to 77.8 in firmness values. Subjective ratings of bulb firmness ranged from 8.5 to 9.0. Depending on the firmness of evaluated breeding lines, our subjective rating system should be adjusted to better distinguish firmness differences between bulbs.

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Robert H. Bors and J. Alan Sullivan

Interspecific crosses with Fragaria moschata (6x) have been hampered by ploidy level differences, poor seed set, and extremely poor seed germination. Modification of pollination practices, embryo rescue, and use of several genotypes has allowed over 80 synthetic tetraploids to be created from 14 cross combinations. Germplasm for the experiment consisted of eight selections of F. moschata (6x), two of F. nubicola (2x), and two of F. viridis (2x). Both 2x × 6x and 6x × 2x crosses were performed. Initially, negligible seed set occurred on F. nubicola and F. viridis when multiple flowers per truss were pollinated. When only one cross was performed per truss, with other flowers removed, seed set was greatly enhanced. F. moschata was much more tolerant of multiple crosses per truss. The crossing combination of F. moschata × F. nubicola gave the worst seed production. Other species combinations were capable of producing good seed set with noticeable differences between individual selections. When achenes were halved, only 1% appeared normal, 2% were underdeveloped or shrunken, the remainder were empty. Many of the malformed and most of the normal embryos germinated using the cut achene method. Achenes were surface-sterilized, cut in half, and placed on MS media with activated charcoal (3g·L–1), sucrose (30g·L–1), and no hormones. Germination occurred only from achenes from fully ripened fruit. Viable hybrids were obtained from 2x × 6x as well as 6x × 2x crosses. Fragaria viridis–F. moschata hybrids closely resembled F. moschata while F. nubicola–F. moschata hybrids were more intermediate in leaf morphology.

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David A. Dierig, Anson E. Thompson, Earl R. Johnson, and Gail H. Dahlquist

Vernonia galamensis is a potential new crop for production of epoxidized oil with many industrial applications. This plant is native to equatorial Africa, and not adapted for culture in temperate zones since it requires a short daylength to initiate flowering and subsequent seed development. One collection of V. galamensis ssp. galamensis var. petitiana, flowered freely and produced seeds during long-day conditions throughout the United States. This variety lacks important plant characters for successful commercialization. The favorable genetic recombination of day-neutral response with more desirable plant growth characteristics, desirable seed oil and fatty acid content from other accessions of V. galamensis has been accomplished in hybrids and segregating populations, and selections are being widely evaluated throughout the U.S..

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Heli Cain Nunez-Grajeda and Sergio Garza-Ortega

Cushaw squash is cultivated in northwest Mexico mainly during the fall and to a lesser degree in the spring season, in which a lack of fruit production in experimental and commercial materials has been observed. This work was done to test 12 lines, 16 hybrids and six landraces regarding fruit and seed weight, flesh color, and soluble solids content (SSC) in both spring and fall seasons in year 2002. Estimates of fruit and seed yield were done. The crop was established by direct seeding at 0.5-m spacing between plants, on both sides of furrow-irrigated beds measuring 15 m long and 4 m wide. In the spring, fruit weight changed from 2.7 to 4.7 kg and seed weight from 17 to 118 g/fruit; fruit yield varied from 3.2 to 38.8 t·ha-1 and seed yield from 18 to 1131 kg·ha-1. Thirty-two percent of the genotypes, including lines and hybrids, but not landraces, were fruitless. SSC and flesh color had values from 4% to 7.5% and from 5.22 to 6.94 Y, respectively. For the fall culture all the genotypes showed good fruit set. Fruit weight in this season changed from 0.8 to 3 kg and seed weight from 22.3 to 97 g/fruit; fruit and seed yield varied from 4 to 28 t·ha-1 and from 135 to 923 kg·ha-1, respectively. All of the landraces were severely infected with squash leaf curl virus and had very low yields. SSC and flesh color, in this season, had values from 3.6% to 10.4% and from 5.1 to 7.94 Y, respectively.

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Mark W. Farnham

Using anther culture to generate doubled-haploid (DH) homozygous lines for use as parents in F1 hybrid crosses has become a common practice in breeding broccoli (Brassica oleracea L. Italica Group). During anther culture and subsequent embryogenesis and plant regeneration, polyploidization of microspore-derived embryos may not occur or it may occur accompanied by a doubling, tripling, quadrupling, octupling, or irregular polyploidization of the genome. Thus regenerants from the process can be haploids, diploids, triploids, tetraploids, octaploids, or aneuploids. The objectives of this research were to 1) conduct repeat cycles of broccoli anther culture using a group of F1 hybrids as anther donors and develop populations of regenerants; 2) analyze resulting populations using DNA flow cytometry and determine the influence of F1 source on frequency of different ploidy levels among regenerants; and 3) compare seed set in broccoli inbreds developed in a traditional selfing program compared to seed set in DH broccoli derived from anther culture. In two cycles (1994 and 1995) of anther culture, anther-derived populations of regenerants were developed using the F1 hybrids `Marathon', `Everest', `High Sierra', and `Futura' as sources of anthers. In 1994, `Everest', `High Sierra', and `Futura' yielded populations that included 2% to 7% haploids, 53% to 56% diploids, 32% to 38% tetraploids, and 5% to 6% other types. `Marathon'-derived regenerants were 5% haploid, 78% diploid, 15% tetraploid, and 2% other, showing significantly more diploids. In 1995, `Marathon' regenerants again included significantly more diploids and fewer tetraploids than those derived from other F1 sources, confirming that the genotype of the anther source affects the frequency of a particular ploidy level among regenerants derived from culture. In manual self-pollinations of 1994 regenerants, only diploids and rare tetraploids set seed. When plants that set no seed were discounted, seed production following manual self pollinations of 1995 regenerants was not significantly different from that of traditional inbreds derived from the same F1 sources.

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Neil O. Anderson and Peter D. Ascher

Chrysanthemum [Dendranthema ×grandiflora Tzvelv. (syn. Chrysanthemum ×morifolium Ramat.)] breeding programs have been selecting for reduced expression of self-incompatibility (via pseudo-self-compatibility) to create inbred families with selected genotypes to serve as parents for F1 hybrid chrysanthemum seed production. However, it is not known to what extent inbreeding is affecting fertility in this outcrossing, heterozygous species. The objective of this research was to assess male/female fertility changes (gain/loss) in successive inbred generations of chrysanthemums. Pseudo-self-compatible chrysanthemum parents (n = 41 inbred, noninbred, and recombinant inbred) were chosen for fertility analyses. As many as three generations of inbreds (I1, I2, and I3) from self-pollinations were created using rapid generation cycling. Female and male fertility levels of the parents and all derived inbred populations were assessed using outcross seed set and pollen stainability, respectively. Average seed set ranges were 0.3% to 96.1% (inbred parents), 24.5% to 38.5% (noninbred parents), and 0.9% to 85.1% (recombinant inbred parents); these began decreasing in the I1 and continued to decline steadily into the I3. Statistically significant (P < 0.05) decreases in seed set occurred in n = 23 (56.1%) inbred families; the remaining inbred families had similar or higher fertility than the parents. Pollen stainability was >50% for the parents, but began declining in some inbred families as inbreeding progressed. Fertility reductions were attributed to inbreeding depression. Lack of significant fertility losses in other inbred families demonstrates the opportunity of selection of fertile inbred parents for use in hybrid seed production.

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Daniel J. Cantliffe

hybrid seed production and plant germplasm conservation. The reader has an opportunity to get information, packed into 796 pages, in almost any area related to seeds. The contributing authors to Dr. Basra's book are excellent and well known for their