There is increasing interest in using methanol and other alcohol fuels as an alternative energy source in the United States and developing nations. However, methanol-fueled vehicles have higher direct emissions of formaldehyde (HCHO) than gasoline-fueled vehicles, which has led to concern about increases in atmospheric concentration of HCHO. Formaldehyde at concentrations of 300, 600, 900, and 1200 μM reduced germination of hydrated Douglas-fir (Pseudotsuga menziesii) pollen in vitro. HCHO concentrations and pH in media containing pollen decreased during the 25-h incubation, with decreases proportional to HCHO concentration. This effect was not seen with heat-killed pollen, which suggests a detoxification mechanism. Ion leakage (measured as electrical conductivity) of pollen increased within 20 h in all HCHO treatments compared to controls. Stress also was indicated by TTC staining, which also decreased after HCHO treatment compared to controls.
A.M. Shirazi and P.S. Muir
Sandra M. Reed
Low seed set has been reported following self-pollinations of flowering dogwood (Cornus florida L.). The objective of this study was to verify the presence of self-incompatibility in C. florida. `Cherokee Princess' stigmas and styles were collected 1, 2, 4, 8, 12, 24, 48, and 72 hours after cross- and self-pollinations, stained with aniline blue and observed using a fluorescence microscope. Pollen germinated freely following self-pollinations, but self-pollen tubes grew slower than those resulting from cross-pollinations. By 48 hours after cross-pollination, pollen tubes had reached the bottom of the style while pollen tubes in self-pollinated flowers had only penetrated the upper third of the style. Evidence of reduced pollen tube growth rate in self-pollinations of `Cherokee Chief' and `Cherokee Brave' was also obtained. This study provides evidence of a gametophytic self-incompatibity system in C. florida. It was also determined that stigmas of C. florida `Cherokee Princess' are receptive to pollen from 1 day prior to anthesis to 1 day after anthesis.
Sylvia J. Brooks and Paul M. Lyrene
Fertility of F1 hybrids and their open-pollinated progeny was studied for the intersectional cross Vaccinium darrowi Camp × V. arboreum Marsh as part of a project to determine the feasibility of using V. arboreum to breed vigorous, drought-tolerant southern highbush blueberry cultivars. The 16 F1 hybrids that were studied were vigorous but very low in fertility. Second generation hybrids [MIKs (mother is known) obtained by open-pollination of the F1s] and MIK derivatives were extremely variable in vigor and fertility, but averaged far higher in fertility than the F1s as evidenced by pollen stainability and amount of pollen produced. F1s produced an average of 0.4 seedlings per 100 pollinated flowers when hand-pollinated in a greenhouse with pollen from V. darrowi, 0.2 when pollinated by V. arboreum and 3.4 when pollinated by cultivated highbush. Some MIKs that were crossed with other MIKs and with cultivated southern highbush were very high in male and female fertility. Female fertility was estimated in greenhouse crosses from fruit set, berry weight, number and weight of seeds, number of plump seeds per berry, and number of seedlings obtained. Male fertility was estimated by pollen stainability with acetocarmine and amount of pollen shed. Chromosome counts showed that three F1s were diploid and that four fertile MIKs were tetraploid. One MIK appeared to be aneuploid. Aneuploidy may explain much of the low fertility found in MIK populations. These results indicate that good progress is being made in returning the hybrid plants to cultivar quality in only a few generations of backcrossing.
Chang-Yeon Yu and John Masiunas
The objective of this study was to investigate the chromosomal and genotypic variation in regenerated plants of Solarium and Lycopersicon. Calli of Lycopersicon peruvianum genotypes PI199380, PI126345, PI251301, and LA1373, along with Solanum ptycanthum were transferred onto media consisting of MS salts with Gamborg vitamins. The shoots formed were rooted in vitro and transferred to greenhouse soil. Actively growing root tips were harvested and pretreated, fixed, hydrolyses and stained. Pollen mother cells were fixed in propionic alcohol solution and stained with aceto-carmine. The number of chromosomes were counted. The greatest variation was in Solanum ptycanthum with chromosome numbers ranging from 18 to 60 (2n=24). Progeny analysis for 12 somaclones of Solarium ptycanthum was done by selfing for two generations. Morphology, shoot height, and weight were determined in each generation. The amount of variation differed among the somaclonal lines.
Sandra M. Reed
The objectives of this study were to evaluate self-incompatibility in Hydrangea paniculata Sieb. and H. quercifolia Bartr. and to determine optimum time for pollination of these two species. Flowers from three genotypes of each species were collected 1, 2, 4, 8, 24, 48, and 72 hours after cross- and self-pollination, stained with aniline blue and observed using a fluorescence microscope. In both species, pollen germination was observed on stigmas of over half of the flowers collected 4 to 72 hours after cross- or self-pollination. Differences in pollen tube length between cross- and self-pollinated flowers were noted from 8 to 72 hours after pollination in H. paniculata and from 24 to 72 hours after pollination in H. quercifolia. By 72 hours after pollination, most self-pollen tubes had only penetrated the top third of the style but cross-pollen tubes had grown to the base of the style and entered 40% to 60% of the ovules. Stigmas of H. paniculata were receptive to pollen from anthesis to 5 days after anthesis, while stigmas of H. quercifolia were receptive from 1 to 5 days after anthesis. This study provides evidence of a gametophytic self-incompatibility system in H. paniculata and H. quercifolia. Occasional self-seed set previously observed in these species was theorized to have been due to pseudo-self compatibility.
Yi He, Hazel Y. Wetzstein, and Barrv A. Palevitz
Fungicides have been shown to negatively affect pollen germination, tube growth, and fruit set in important crops. However, little is known regarding possible modes of action in higher plant cells. To address this, the effects of propiconazole or benomyl on pollen germination and tube growth were evaluated in Tradescantia virginiana using light microscopy and immunocytochemistry. Concentrations were selected at levels that had inhibitory effects, but did not totally arrest germination and tube elongation, i.e., propiconazole and benomyl were added at 0, 102, 136, or 170 μl·liter–1, and 0, 480, 600, or 720 mg·liter–1, respectively. Both fungicides inhibited germination, cytoplasmic streaming, tube elongation, and induced abnormal tube morphology and cytoskeletal distribution. Propiconazole-treated tubes had weaker microfilament signals, with amorphous staining. Microtubule (Mt) distribution was severely affected. In benomyl-treated tubes, Mts were fewer in number, fragmented, sinuous, and increasingly disorganized. Possible mechanism(s) will be discussed.
Sandra M. Reed
The objectives of this study were to evaluate self-fertility and to determine the effectiveness of pollinations made over a 4-day period in Japanese snowbell, S. japonicum Sieb. & Zucc. Pollen germination and pollen tube growth were observed in stained styles following cross- and self-pollinations made from 1 day before to 2 days after anthesis. One month after pollination, fruit set averaged 40% in cross-pollinations and 14% in self-pollinations. Two months later, about one-third of the fruit resulting from cross-pollinations had aborted and only one fruit remained from the self-pollinations. This study demonstrated that stigmas of S. japonicum are receptive for at least 4 days and that flowers should be emasculated prior to making controlled cross-pollinations.
Thomas H. Boyle
True-breeding lines of Zinnia marylandica Spooner, Stimart and Boyle [allotetraploids of Z. angustifolia H.B.K. and Z. violacea Cav. (2n = 46)] were reciprocally backcrossed with diploid and autotetraploid forms of Z. angustifolia (2n =22 or 44) and Z. violacea (2n =24 or 48). In most cases, backcrosses were more successful with Z. angustifolia and Z. violacea as autotetraploids than as diploids. Seed-generated, backcross (BC1) families were obtained by crossing Z. marylandica (as female) with autotetraploid Z. angustifolia or autotetraploid Z. violacea. BC1 plants were phenotypically intermediate between the two parental lines for most morphological characters. Crosses between Z. marylandica and autotetraploid Z. angustifolia yielded BC1 plants with 33% stainable pollen, whereas crosses between Z. marylandica and autotetraploid Z. violacea yielded BC1 plants that produced malformed, poorly-stained pollen. No embryos were observed in capitula collected from field-grown BC1 plants. BC1 hybrids of Z. marylandica and autotetraploid Z. violacea produced larger capitula and more ray florets than Z. marylandica, and exhibited novel combinations of floral pigments not observed in Z. marylandica ray florets. BC1 hybrids of Z. marylandica and Z. violacea have commercial potential as seed-propagated, bedding plants.
Michele A. Stanton, Joseph C. Scheerens, Richard C. Funt, and John R. Clark
We investigated the response of staminate and pistillate floral components of Prime-Jan™ and Prime-Jim™ primocane-fruiting blackberry (Rubus L. subgenus Rubus Watson) to three different growth chamber temperature regimes, 35.0/23.9 °C (HT), 29.4/18.3 °C (MT), and 23.9/12.8 °C (LT). Temperature was negatively related to flower size and morphological abnormalities in floral structures were evident in 41% and 98% of the MT- and HT-grown plants, respectively. The viability (stainability) of pollen from LT- and MT-grown Prime-Jan™ flowers exceeded 70%; that of Prime-Jim™ pollen was significantly reduced (<40%) by the MT regime. Pollen in-vitro germinability was negatively influenced by temperature but was unaffected by cultivar. LT-grown pollen held at 23.9 °C retained 63% of its original germinability over a 32-hour period; the germinability of LT-grown pollen held at 35.0 °C was decreased by 97% from its original level after 16 hours. Virtually all flowers cultured under HT conditions were male-sterile, exhibiting structural and/or sporogenous class abnormalities including petaloidy, malformation of tapetal cells, and microspores or failure of dehiscence. The duration of stigma receptivity, pistil density, and drupelet set were also negatively influenced by increasing temperature; values for these parameters of floral competency among control plants were reduced by 51%, 39%, and 76%, respectively, in flowers cultured under HT conditions. Herein, flowering and fruiting parameters and presumably the yield potential of Prime-Jan™ and Prime-Jim™ were adversely affected by increased temperature. However, assessment of their adaptative response to heat stress under field conditions awaits experimentation.
Jon T. Lindstrom, Chih-Hsien Lei, Michelle L. Jones, and William R. Woodson
Mature pollen from Petunia hybrida contains significant levels of 1-aminocyclopropane-1-carboxylic acid (ACC), and this ACC is thought to play a role in pollination-induced ethylene by the pistil. We investigated the developmental accumulation of ACC in anthers and pollen. The level of ACC in anthers was very low until the day before anthesis, at which time it increased 100-fold. A 1.1-kb partial ACC synthase cDNA clone (pPHACS2) was amplified from total RNA isolated from mature anthers by reverse transcriptase, followed by polymerase chain reaction using oligonucleotide primers synthesized to conserved amino acid sequences in ACC synthases. The expression of pPHACS2 mRNA during anther development was correlated with the accumulation of ACC and was localized to the pollen grain. The pPHACS2 cDNA was used to identify the PH-ACS2 gene from a library of genomic DNA fragments from Petunia hybrida. PH-ACS2 encoded an ACC synthase transcript of four exons interrupted by three introns. The ACC synthase protein encoded by the PH-ACS2 gene shared >80% homology with ACC synthases from tomato (LE-ACS3) and potato (ST-ACS1a). A chimeric PH-ACS2 promoter-β-glucuronidase (GUS) gene was used to transform petunia and transgenic plants were analyzed for GUS activity. GUS staining was localized to mature pollen grains and was not detected in other tissues. Despite similarities to LE-ACS3, we did not detect GUS activity under conditions of anaerobic stress or in response to auxin. A series of 5-prime-flanking DNA deletions revealed that sequences within the PH-ACS2 promoter were responsible for pollen-specific expression.