Male Clones Research Articles

A chromosome-level genome of the human blood fluke Schistosoma japonicum identifies the genomic basis of host-switching.

The evolution and adaptation of S.japonicum, a zoonotic parasite that causes human schistosomiasis, remain unclear because of the lack of whole-genome data. We construct a chromosome-level S.japonicum genome and analyze it together with 72 samples representing six populations of the entire endemic region. We observe a Taiwan zoophilic lineage splitting from zoonotic populations ∼45,000 years ago, consistent with the divergent history of their intermediate hosts. Interestingly, we detect a severe population bottleneck in S.japonicum, largely coinciding with human history in Asia during the last glacial maximum. We identify several genomic regions underlying natural selection, including GATAD2A and Lmln, both showing remarkable differentiation among different areas. RNAi knockdown suggests association of GATAD2A with parasite development and infection in definitive hosts, while Lmln relates to the specificity of the intermediate hosts. Our study provides insights into the evolution of S.japonicum and serves as a resource for further studies.

Elemental composition in male and female genotypes of betel vine (Piper betle L.)

Betel vine (Piper betle L.,) is an evergreen perennial, dioecious creeper belongs to the family Piperaceae and cultivated commercially in South East Asian countries. In present study the sexually dimorphic genotypes were evaluated to know their elemental composition (nitrogen, phosphorous, potassium, iron, zinc, copper and manganese) and found significant results within male and female clones. On the other side, except copper the gender differences were not found significant. The nitrogen content was higher among major elements in Sirugamani-1 (6.63%). The phosphorous concentration was higher in genotype CARI-6 (0.72%). Similarly, potassium levels was found higher in genotype Kapoori TN (2.83%). The trace element copper was higher in male genotypes Kapoori TN and Swarna Kapoori (36.21 mg/kg). The copper concentration was slightly higher in male clones over female counterparts. In other hand iron was found higher among all the trace elements and higher contents in genotype Kapoori Chintalapudi (84.38 mg/kg).

Variation and Genetic Parameters of Leaf Morphological Traits of Eight Families from Populus simonii × P. nigra

Leaf morphology in Populus L. varies extensively among sections, species and clones under strong genetic control. P. nigra L. (section Aigeiros), with large and triangular leaves, is a commercial forest tree of economic importance for fast growth and high yield in Europe. P. simonii Carr. (section Tacamahaca) with small land rhomboid ovate leaves performs cold and dry resistance/tolerance in the semi-arid region of Northern China. Leaf morphological traits could be used as early indicators to improve the efficiency of selection. In order to investigate the genetic variation pattern of leaf morphology traits, estimate breeding values (combining ability), as well as evaluate crossing combinations of parents, 1872 intersectional progenies from eight families (P. simonii × P. nigra) and their parents were planted with cuttings for the clonal replicate field trial in Northern China. Four leaf size traits (area, perimeter, length, width) and roundness were measured with leaf samples from the 1-year-old clonal plantation. Significant differences regarding leaf traits were found between and among three female clones of P. simonii from Inner Mongolia, China and six male clones of P. nigra from Casale Monferrato, Italy. The genetic variation coefficient, heritability and genetic variance component of most traits in male parents were greater than these of female parents. Heritability estimates of male and female parents were above 0.56 and 0.17, respectively. Plentiful leaf variations with normal and continuous distributions exited in the hybrid progenies among and within families with the genetic variation coefficient and heritability above 28.49 and 0.24, respectively. Heritability estimates showed that leaf area was the most heritable trait, followed by leaf width. The breeding value ranking of parents allowed us to select the parental clones for new crosses and extend the mating design. Two male parental clones (N430 and N429) had greater breeding values (general combining ability, GCA) of leaf size traits than other clones. The special combining ability (SCA) of the crossing combination between P. simonii cl. ZL-3 and P. nigra cl. N430 was greater than that of others. Eight putatively superior genotypes, most combined with the female parental clone ZL-3, can be selected for future testing under near-commercial conditions. Significant genetic and phenotypic correlations were found between five leaf morphology traits with the coefficients above 0.9, except for leaf roundness. The results showed that leaf morphology traits were under strong genetic control and the parental clones with high GCA and SCA effects could be utilized in heterosis breeding, which will provide a starting point for devising a new selection strategy of parents and progenies.

Estimation of chlorophyll contents in male and female genotypes of betel vine (Piper betle L.)

Betel vine [Piper betle (L)] is valued for its leaves due to its masticatory health benefits. The cultivars in betelvine are mostly the land races which are cultivated locally. The present study aimed to estimate the chlorophyll contents among male and female genotypes collected from different parts of India. The chlorophyll a, b and total chlorophyll contents among male and female clones varied significantly within genders. The females had slightly higher contents of chlorophyll over male clones. The highest total chlorophyll recorded in genotype Hirehalli Local (2.363 mg/g) followed by, IIHR BV 67 (2.082 mg/g) in females. Among male clones Shirpurkata (2.238 mg/g) followed by, clone Kapoori TN (2.180 mg/g) had higher total chlorophyll contents. The general observation prevails that, male leaves has slightly lesser in chlorophyll contents over females. Present study could able to identify the male genotypes (Shirpurkata and Kapoori TN) which are comparable in chlorophyll contents over female clones.

Sex-specific responses of Populus deltoides to combined salinity and calcium under waterlogging conditions

In this study, we investigated the sex-specific ion uptake and physiological and biochemical responses to combined salinity and calcium treatments of male and female Populus deltoides under waterlogging conditions. Results indicate that both P. deltoides clones were sensitive to salinity and calcium. Under well-watered conditions, salinity stress could especially increase the Na content in female P. deltoides clones, whereas salinity and calcium could increase the K and Ca content in male clones. Waterlogging could evidently stimulate Na content in leaves under salinity, especially in female P. deltoides clones. However, waterlogging had no visible effects on the amount of Ca absorption between the two clones under abundant calcium conditions. The physiological and biochemical responses to combined salinity and calcium exceeded the effects of salinity or calcium individually. However, no significant difference was observed, hence indicating the similar responses of male and female P. deltoides clones. The lower Na content and Na:Ca ratio, and the higher K and Ca content in male clones suggest that the male clones could maintain ion homeostasis better than the female clones. The sex-specific differences in net photosynthetic rate, intercellular CO2 concentration, effective quantum yield of photosystem II, photochemical quenching coefficient, non-photochemical quenching coefficient, photosynthetic electron transport rate, content of superoxide anions, hydroxyl radicals, H2O2, soluble protein, and activities of superoxide dismutase under certain conditions suggest that female P. deltoides clones are more sensitive to salinity, calcium, and the combination of both than their male counterparts.

Assessments of growth performance, crown structure, stem form and wood property of introduced poplar clones: Results from a long-term field experiment at a lowland site

Climate change brings about more frequent short-term waterlogging leading to unfavorable conditions for tree growth. Poplar is a preferred tree species for fast-growing plantations. Although limited short-term comparative studies exist with poplars that examine clonal variation in growth and physiological responses to waterlogging, a comprehensive assessment on growth performance, crown structure and wood property of poplar clones is lacking from a long-term study. This study compares the clonal variations in survival rate, growth, crown structure, stem form and wood property of poplar clones to better match the genotype with specific site. In line with the result of the preliminary evaluation, 17 poplar clones were harvested to further investigate crown structure, stem form and wood property after 24 growing seasons. A significant difference in tree growth was observed among the clones, ranging from 1.08 to 2.64 m3 for tree volume, whereas male clones generally had a better growth performance than the female. Hierarchical cluster analysis indicated the 17 poplar clones can be classified into three distinct groups in term of mean values of branching angle, branch numbers and branch base diameter. There existed visible differences in breast-height form factor and height-diameter ratio among the poplar clones, and four distinct groups can be divided based on Euclidean distance of stem form indexes. Significant clonal variations in wood basic density (WD) and microfibril angle (MFA) were observed among the sampled clones, and the mean clonal WD and MFA at breast height ranged from 0.325 to 0.390 g/cm3 and 11.8 to 16.7° respectively. Based on the TOPSIS method, a comprehensive assessment of adaptability and prioritization of the 17 clones was made from tree volume growth, stem form, branching traits and wood density, and some superior poplar clones were recommended for future applications in the similar lowland sites.

Clonal Variation in Growth Response of Populus deltoides Bartr. to Nitrogen Fertilization Under Nursery Conditions

Populus deltoides, an extensively planted poplar in Jammu and Kashmir and is one of the ten most prominent tree species grown in the State. A strapping clone x fertilizerinteraction was observed fordiameter, height and biomass. Growth response of six male clones of this species to three levels ofnitrogen (50, 75 and100 kg ha 1 ) was evaluated in nursery for two years to produce healthy and vigorous nursery stock. While application of N- fertilizer @ 75 kg ha 1 recorded maximum height, diameter, number of branches plant', numberofleaves plant', leafarea plant', internodal length and total biomass (fresh and dry) plant', the application of 100 kg N ha ' was observed to exert toxic effects and reduced the growth performance. In terms ofefficient utilization ofNitrogen fertilizer (NUE), the application of 50 kg N ha ' yield maximum biomass p er unit of N- used by the plant. The male clones have recently received a lot of attention due to the increasing cotton mince produced by female clones in Kashmir valley. In currentscenario male clones are the betterperspective to replace the female ones without compromising the economy generated by this species.

WYNIKI BADAŃ NAD MIESZAŃCEM TOPOLI POPULUS MAXIMOWICZII X POPULUS SIMONII ‘FASTIGIATA’ W NADLEŚNICTWIE KALISKA

The article describes the research carried out in the Kaliska Forest District on a new variety of poplar ‘Zbigniew Stecki’. This poplar is an attractive hybrid of Populus maximowiczii x Populus simonii 'Fastigiata', occurring as a columnar type male clone. The influence of rainfall and temperature on the annual and total tree height was investigated. The amount of rainfall during the growing season does not have a significant influence on the shoot growth in the poplar variety Zbigniew Stecki.

In vitro studies on male clone of Riccia discolor--Effects of sugars

In vitro studies on male clone of Riccia discolor--Effects of sugars

The ideal auricular protrusion – An interactive perceptual pilot study

PurposeExcessive ear protrusion (prominauris) can negatively affect facial appearance. Because the concept of an ideal auriculocephalic angle is controversial, however, it is difficult to define when an obtrusive auricle requires surgical intervention. It is often assumed that angles exceeding 30° require corrective surgery. However, little is known about public perception of ear protrusion. This study aimed to assess perceptions of different degrees of auricular protrusion. Materials and methodsWe conducted an interactive panel survey. Male and female evaluators assessed digitally processed cloned images of a male and female model that depicted various ear protrusions ranging from 0° to 90°. Predictor variables were the sex of the evaluator, the sex of the clone and the extent of auricular protrusion. The outcome variable was the overall attractiveness of auricular appearance. ResultsForty-four evaluators (students of dental medicine, mean age 25.43 years) participated in the study: 22 women and 22 men. The study results revealed sexual dimorphism, with the attractiveness of corresponding male and female clones rated differently. Male evaluators favoured female ear positions that fitted closer to the head; preferred alignments for female clones were: 18° (Mean) ± 9.38° (SD), p = 0.006; for male clones: 30° ± 7.94°, p = 0.003. Ear protrusion starts to negatively affect aesthetic appearance for male evaluators at positions of 42° ± 4.87° (female clone) and 54° ± 8.22° (male clone), p = 0.001. The panel of evaluators agreed to perceive angles of 21–24° ± 8.10° (female clones), ± 8.33° (male clones), as the ideal degree of protrusion in both sexes (p = 0.158). ConclusionIn this study, perception of ear appearance depended on the sex of both the evaluator and the clone being assessed. Obtrusive ears were more readily accepted in male faces than in female ones. Nevertheless, the overall panel perception shows a preference for rather close fitting, unobtrusive ear alignments. For planning of corrective ear surgery, a target angle of approximately 22° can be considered as a desirable result.

Development of cardoon male sterile clones

Development of cardoon male sterile clones

Roots play a key role in drought-tolerance of poplars as suggested by reciprocal grafting between male and female clones

Drought stress influences the growth of plants and thus grafting has been widely used to improve tolerance to abiotic stresses. Poplars possess sex-specific responses to drought stress, but how male or female rootstock affect the grafted plant is little known. To explore the mechanisms underlying changes in drought tolerance caused by grafting, we investigated the changes in growth, leaf traits, gas exchange and antioxidant enzyme activities of reciprocally grafted seedlings between Populus euramericana cv. "Nanlin895" (NL-895) (female) and Populus deltiodes cv."3412" (NL-3412) (male) under water deficit stress with 30% field capacity for 30 d. Results showed that drought stress affected adversely growth, morphological, and physiological characteristics in all seedlings studied. Grafted seedlings with male roots can effectively alleviated the inhibition of growth induced by drought stress, as shown by higher WUE, activities of SOD, POD and CAT, and lower levels of lipid peroxidation. Male seedlings with female roots were found to be less tolerance to drought than non-grafted male clones and female scions with male roots, but more tolerance than non-grafted female clones. This results suggested that drought tolerance of grafted seedlings is primarily caused by the rootstock, although the scion also affects the grafted plant. Thus, paying attention on the root genotype can provide an important means of improving the drought tolerance of poplars.

Association between metabolic resistances to atrazine and mesotrione in a multiple-resistant waterhemp (Amaranthus tuberculatus) population

AbstractMetabolic resistances to atrazine (atz-R) and mesotrione (meso-R) occur in several waterhemp [Amaranthus tuberculatus (Moq.) Sauer] populations in the United States. Interestingly, although metabolic atz-R but mesotrione-sensitive A. tuberculatus populations have been reported, an Amaranthus population has not been confirmed as meso-R but atrazine-sensitive, implying an association between these traits. Experiments were designed to investigate whether the single gene conferring metabolic atz-R plays a role in meso-R. An F2 population was generated from a multiple herbicide–resistant A. tuberculatus population from McLean County, IL (MCR). A cross was made between a known meso-R male clone (MCR-6) and a herbicide-sensitive female clone from Wayne County, IL (WCS-2) to develop an F1 population. Survival of MCR-6 plants following atrazine POST treatment (14.4 kg ha−1) indicated the male parent was homozygous atz-R. F1 plants were intermated to obtain a segregating pseudo-F2 population. Dose–response and metabolic studies conducted with mesotrione using F1 plants indicated intermediate biomass reductions and metabolic rates compared with MCR-6 and WCS. F2 plants were initially treated with either mesotrione (260 g ha−1) or atrazine (2 kg ha−1) POST, and after 21 d of recovery, vegetative clones from surviving resistant plants were subsequently treated with the other herbicide. When mesotrione was applied first, the meso-R frequency was 8.2%, and when atrazine was applied first, the atz-R frequency was 75%. However, the meso-R frequency increased to 16.5% following preselection for atz-R, and 100% of surviving meso-R plants were atz-R. Our findings indicate that the gene conferring metabolic atz-R is also involved with the meso-R trait within the population tested.

WOMEN IN THE IDENTITY CRISIS OF FEMINISM; A CRITICAL ANALYSIS ON GENDER MOVEMENT BASED ON ISLAMIC PSYCHOLOGY PERSPECTIVE

<p><em>This paper attempted to analyze critically the women's movement that had been developing so far and tried to offer the ideology of the gender movement based on the values </em><em>of humanism of Islamic Psychology. This study used a qualitative methodology with a literature approach. The results showed that (1) the male clone tendency in the women's movement had actually been predicted by various parties, including by the female activists. Because of the obsession to equalize the active role of women as equal to men, the women's movement was trapped in standardizing themselves with male masculinity figures. On one hand, they rejected male domination on women, but on the other hand the ambitions of their movements are directed towards seizing the dominant patriarchal system and violating it under the authority of women. Even though Islam places women in the frame of proportionally optimistic rational roles. That is, Islam does not make women fully pretend to be 'backward' entities so that it does not allow them to gain enlightenment and Islam rejects thoughts that are too optimistic to position 'front' women as the sole determinant of their lives and must be above men. Both of these views have reduced women to fall into the destruction or glorification of human quality by ignoring the greatness and power of Sunnatullah over the surrounding conditions; (2) Acording to the values </em><em>of humanism of Islamic Psychology, the ideology of the gender movement should depart from the needs and be based on a) Women’s self-actualization rather than self-exploitation; b) Women’s Active participation rather than their mobilization and domestication. (c) Partnership rather than rivalism.</em></p>

Efficient generation of GHR knockout Bama minipig fibroblast cells using CRISPR/Cas9-mediated gene editing.

Dwarfism, also known as growth hormone deficiency (GHD), is a disease caused by genetic mutations that result in either a lack of growth hormone or insufficient secretion of growth hormone, resulting in a person's inability to grow normally. In the past, many studies focusing on GHD have made use of models of other diseases such as metabolic or infectious diseases. A viable GHD specific model system has not been used previously, thus limiting the interpretation of GHD results. The Bama minipig is unique to Guangxi province and has strong adaptability and disease resistance, and an incredibly short stature, which is especially important for the study of GHD. In addition, studies of GHR knockout Bama minipigs and GHR knockout Bama minipig fibroblast cells generated using CRISPR/Cas9 have not been previously reported. Therefore, the Bama minipig was selected as an animal model and as a tool for the study of GHD in this work. In this study, a Cas9 plasmid with sgRNA targeting the first exon of the GHR gene was transfected into Bama minipig kidney fibroblast cells to generate 22 GHR knockout Bama minipig kidney fibroblast cell lines (12 male monoclonal cells and 10 female monoclonal cells). After culture and identification, 11 of the 12 male clone cell lines showed double allele mutations, and the rate of positive alteration of GHR was 91.67%. Diallelic mutation of the target sequence occurred in 10 female clonal cell lines, with an effective positive mutation rate of 100%. Our experimental results not only showed that CRISPR/Cas9 could efficiently be used for gene editing in Bama minipig cells but also identified a highly efficient target site for the generation of a GHR knockout in other porcine models. Thus, the generation of GHR knockout male and female Bama fibroblast cells could lay a foundation for the birth of a future dwarfism model pig. We anticipate that the "mini" Bama minipig will be of improved use for biomedical and agricultural scientific research and for furthering our understanding of the genetic underpinnings of GHD.

A battle of the sexes? Sex ratio and performance at small scales in dioecious Mercurialis perennis

Mercurialis perennis is a dioecious clonal woodland herb. Its male-biased shoot sex ratio may result from differential selection (with skewed genet sex ratio) and/or differential performance (fairly even genet sex ratio but skewed shoot sex ratio since male clones grow more vigorously or have a higher flowering propensity). To test both scenarios, a total of 1024 shoots were mapped and excavated in five 1 m2 plots in an old-growth forest in northeastern Germany. SSR genotyping revealed a less skewed intrinsic male:female shoot sex ratio (1.6) and a nearly even genet sex ratio. The sexes showed intermingling rather than sexual niche segregation. Most of the 36 genets found split into numerous independent units (merigenets). A slight sexual size dimorphism (SSD) was found: male clones possess on average more shoots, have longer stolons and a higher proportion of flowering shoots. Analyses of resource allocation patterns showed 1.5–2.8 times higher costs for sexual reproduction (dry mass, nitrogen) for females. Males invest surplus resources into leaves and stolons, accentuating the SSD; although for genets the difference may be smaller due to the higher flowering rate of males (53% vs. 34%). Photosynthetic performance, deduced from rapid light curves measured with a portable fluorometer, was similar for both sexes, especially in spring; the main challenge being the transformation from light to shade plant with tree foliation. Our data indicate that differential performance accounts for the male bias in shoot sex ratio; competition between sexes is unlikely given the strongly intermingled clones.

Mass killing by female soldier larvae is adaptive for the killed male larvae in a polyembryonic wasp

Self-sacrifice is very rare among organisms. Here, we report a new and astonishing case of adaptive self-sacrifice in a polyembryonic parasitic wasp, Copidosoma floridanum. This wasp is unique in terms of its larval cloning and soldier larvae. Male clone larvae have been found to be killed by female soldier larvae, which suggests intersexual conflict between male and female larvae. However, we show here that mass killing is adaptive to all the killed males as well as the female soldiers that have conducted the killing because the killing increases their indirect fitness by promoting the reproduction of their clone sibs. We construct a simple model that shows that the optimal number of surviving males for both male and female larvae is very small but not zero. We then compare this prediction with the field data. These data agree quite well with the model predictions, showing an optimal killing rate of approximately 94–98% of the males in a mixed brood. The underlying mechanism of this mass kill is almost identical to the local competition for mates that occurs in other wasp species. The maternal control of the sex ratio during oviposition, which is well known in other hymenopterans, is impossible in this polyembryonic wasp. Thus, this mass kill is necessary to maximize the fitness of the female killers and male victims, which can be seen as an analogy of programmed cell death in multicellular organisms.

Negative correlation between ash dieback susceptibility and reproductive success: good news for European ash forests

European ash ( Fraxinus excelsior L.) trees with low susceptibility to ash dieback have higher reproductive fitness compared to highly susceptible trees, although most pronounced for female success. Selection at generation turnover therefore supports the future recovery of ash forests. The introduced invasive pathogen Hymenoscyphus fraxineus (T. Kowalski) Baral, Queloz, and Hosoya cause extensive damage on European ash (Fraxinus excelsior L.). Heritable variation in susceptibility to ash dieback has been observed among ash trees in natural and planted populations, but it is not clear how variation in susceptibility influences reproductive fitness. We hypothesize that healthier male and female trees contribute more gametes to the following generation compared to unhealthy ones. We tested the hypothesis by studying gender, seed production, and paternal success in a clonal field trial with 39 replicated clones. In the trial, the susceptibility level of each clone has been recorded in terms of percent crown damage since 2007. We used a linear regression model to explore the relationship between susceptibility and reproductive success (female and male). The clones revealed a clear gender dimorphism with an approximate 2:2:1 male/female/hermaphrodite ratio. Females with low levels of crown damage produced substantially more seeds compared to highly damaged females. The male clone with the lowest level of susceptibility was the most effective pollen donor, but highly susceptible males also sired some offspring. The results overall represent good news for the potential recovery of ash forests: selection against most susceptible genotypes at generation turnover is expected to facilitate building up disease resistance in ash populations.

Development of anthracnose resistant hybrids of the Greater Yam (Dioscorea alata L.) and interspecific hybrids with D. nummularia Lam.

Controlled pollinations were conducted to develop Dioscorea alata L. hybrids resistant to anthracnose (Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. Overall, 10,410 hand pollinations were carried out with Indian (IN) and Vanuatu (VU) clones in 53 different parental combinations, including diploids (2x) and tetraploids (4x) female and male plants. Pollination efficiency and seed set were comparable for IN2x × VU2x (54.6% and 23.0%) and IN2x × IN2x (57.4% and 28.9%). However, seedling survival with better resistance to anthracnose in nursery under natural disease pressure was higher for IN2x × IN2x (67.3%) compared to IN2x × VU2x (47.8%). IN2x female clones were crossed with VU2x and VU4x male clones in 12 parental combinations each. Pollination efficiency was higher for 2x parents (54.6%) compared to 4x parents (47.0%) but seed set (32.8%) and seedling survival (71.7%) were significantly higher for crosses between IN2x × VU4x. Overall, remarkable variation in pollination efficiency, seed set and seedling survival was observed between different parental combinations even when the same pollen parent was used. Interspecific hybridization between D. alata IN2x females and D. nummularia Lam. Vanuatu polyploid male was found to be successful with considerable variation in pollination efficiency, seed set and seedling survival (49.0, 20.8 and 35.3%). The hybrids exhibited intermediate characters between the two species with high tuber yield, anthracnose resistance but with fast tuber flesh oxidation. These results demonstrate the scope of polyploid and interspecific hybridization and the need for the international exchange of germplasm for D. alata genetic improvement.

Phenotypic Expression and Stability in a Large-Scale Field Study of Genetically Engineered Poplars Containing Sexual Containment Transgenes.

Genetic engineering (GE) has the potential to help meet demand for forest products and ecological services. However, high research and development costs, market restrictions, and regulatory obstacles to performing field tests have severely limited the extent and duration of field research. There is a notable paucity of field studies of flowering GE trees due to the time frame required and regulatory constraints. Here we summarize our findings from field testing over 3,300 GE poplar trees and 948 transformation events in a single, 3.6 hectare field trial for seven growing seasons; this trial appears to be the largest field-based scientific study of GE forest trees in the world. The goal was to assess a diversity of approaches for obtaining bisexual sterility by modifying RNA expression or protein function of floral regulatory genes, including LEAFY, AGAMOUS, APETALA1, SHORT VEGETATIVE PHASE, and FLOWERING LOCUS T. Two female and one male clone were transformed with up to 23 different genetic constructs designed to obtain sterile flowers or delay onset of flowering. To prevent gene flow by pollen and facilitate regulatory approval, the test genotypes chosen were incompatible with native poplars in the area. We monitored tree survival, growth, floral onset, floral abundance, pollen production, seed formation and seed viability. Tree survival was above 95%, and variation in site conditions generally had a larger impact on vegetative performance and onset of flowering than did genetic constructs. Floral traits, when modified, were stable over three to five flowering seasons, and we successfully identified RNAi or overexpression constructs that either postponed floral onset or led to sterile flowers. There was an absence of detectable somaclonal variation; no trees were identified that showed vegetative or floral modifications that did not appear to be related to the transgene added. Surveys for seedling and sucker establishment both within and around the plantation identified small numbers of vegetative shoots (root sprouts) but no seedlings, indicative of a lack of establishment of trees via seeds in the area. Overall, this long term study showed that GE containment traits can be obtained which are effective, stable, and not associated with vegetative abnormalities or somaclonal variation.