Leaf surface factors of transgenic Bt cotton associated with the feeding behaviors of cotton aphids: A case study on non-target effects

The present paper reports case study results of the risk assessment of transgenic Bt cotton on a non-target pest, cotton aphid, Aphis gossypii. Several types of techniques, i.e., electrical penetration graph (EPG), light and electron microscopy, bioassays and chemical analysis, were applied to investigate physical and chemical leaf factors of 2 transgenic Bt cotton lines (GK12 and GK19) and their pa-rental non-Bt cotton line (Simian3) associated with searching and feeding behaviors of cotton aphids on leaves or leaf extracts of cotton plants. EPG results showed that there were some differences among behaviors of cotton aphids on 2 Bt cotton and 1 non-Bt cotton lines. Cotton aphids performed similarly to leaf surface extracts from 3 cotton lines; and leaf surface chemicals, mainly volatiles and waxes, were almost identical in the components and concentrations among the cotton lines. However, three cotton lines were quite different from each other in the densities of certain kinds of covering trichomes. Therefore, the relationships between the physical characteristics and the searching behaviors of cotton aphids on the three cotton lines were constructed as the regression equations. Glandular trichomes and covering trichomes with 5 branches influenced the cotton aphids' searching behaviors effectively; and other trichomes with other branches affected aphids in varying ways. These results demonstrated that leaf surface physical factors of transgenic Bt cotton lines different from their parental non-Bt line could affect the penetration behaviors of non-target cotton aphids. Cotton aphids penetrate and feed more easily on two Bt cotton lines than on the non-Bt cotton line.     

浙江省植被覆盖时空动态及其与生态气候指标的关系

对1982-2006年间浙江省归一化植被指数（NDVI）的时空变化及其与关键生态气候指标（生物热量指数和生物干湿度指数）的相关性进行了研究.结果表明：研究期间,浙江省植被覆盖总体呈缓慢下降趋势,NDVI显著减少的地区约占全省面积的30.71%;生态气候指标与滞后一年的NDVI显著相关,其中生物热量指数与NDVI呈显著负相关,生物干湿度指数与NDVI呈显著正相关,说明研究区湿度的增加可促进植被长势,而过高的热量对植被生长则具有明显的抑制作用.     

The Development and a Cytogenetic Study of Monosomics of Gossypium Hirsutum L.

Monosomics of cotton (Gossypium hirsutum L.) were obtained by irradiation of pollen by -rays and by irradiation of seeds by thermal neutrons. Many monosomics were derived directly from irradiation, but a number of monosomics were also recovered in the progeny of plants with translocations and of desynaptic plants. Only 28 primary monosomics showed normal pairing at metaphase-1 of meiosis. The others formec rare trivalents or additional univalents. Partial desynapsis was detected in some monosomics. The pollen fertility levels of monosomics are presented. New morphological characters were detected among the monosome plants of cotton.     

北疆高产棉花密度与打顶时序调控的生态位适宜度研究

以北疆棉区高产 (2 0 0 0kg·hm-2 )棉花 (G .hirsutumL .)为研究对象 ,引进生态位适宜度理论对受密度和打顶时序调控的高产棉花生态位适宜度进行了研究。经果表明 ,在密度梯度上 ,随密度增加棉花生态位适宜度值增加。 1 5 0× 1 0 3 株·hm-2 密度下 ,分次打顶降低了生态位适宜度 ,导致减产 ;在高密度种植条件下 ,分期打顶能提高生态位适宜度 ,有利于提高产量。在 2 2 5× 1 0 3 和 30 0× 1 0 3 株·hm-2 密度条件下 ,与同密度下 1次打顶 (7/5 )相比 ,2次打顶增产 4 5 %和 1 1 3%,3次打顶增产 7 5 %和 5 9%。 1 5 0× 1 0 3 株·hm-2 密度下 ,棉田苗蕾期LAI较小、漏光损失大 ,光合势低 ,是实现超高产的主要限制因子 ,花铃期的分期打顶导致减产。在 2 2 5× 1 0 3 和 30 0× 1 0 3 株·hm-2 密度条件下 ,棉花苗蕾期LAI增加 ,分期打顶保证了花铃期较适宜的LAI ,提高了棉花生态位适宜度 ,有利于提高产量 ;1次打顶处理 (7月 5日 )比本区生产上的打顶时间 (7月 1 0日左右 )提早了 5d左右 ,使高密度棉花群体LAI的发展受到了一定程度地限制 ,避免了因密度高群体透光条件的恶化而降低了其生态位适宜度值 ,造成减产损失。     

陆地棉核不育株扦插繁殖与宿生保持及杂种优势利用

利用广西南宁冬季无霜冻的气候特点,对陆地棉(Gossypium hirsutum L.)洞A细胞核雄性不育扦插株与宿生株及其杂交一代进行比较研究.结果表明:(1)1 a生扦插株开花较早,茎粗、主茎节间长度、铃重、子指4个性状显著好于实生株,但扦插株的种子产量显著低于实生株,其原因是果枝扦插形成的僵苗占扦插株的23.04%,而且扦插株全为不育株,实生株中则有50%左右的可育株;(2)2 a生扦插株性状与实生株无显著差异;(3)由扦插繁殖的不育株和种子繁殖的不育株配制的杂交F1代的产量与纤维品质无显著差异.因此认为,扦插繁殖陆地棉核不育株用于多年杂交制种是可行的,选择营养枝扦插是陆地棉核不育株扦插繁殖需要注意的关键问题. 宿生株及其杂交一代进行比较研究.结果表明:(1)1 a生扦插株开花较早,茎粗、主茎节间长度、铃重、子指4个性状显著好于实生株,但扦插株的种子产量显著低于实生株,其原因是果枝扦插形成的僵苗占扦插株的23.04%,而且扦插株全为不育株,实生株中则有50%左右的可育株;(2)2 a生扦插株性状与实生株无显著差异;(3)由扦插繁殖的不育株和种子繁殖的不育株配制的杂交F1代的产量与纤维品质无显著差异.因此认 ,扦插繁殖陆地棉核不育株用于多年杂交制种是可行的,选择营养枝扦插是陆地棉核不育株扦插繁殖需     

Antimicrobial terpenoids of Gossypium: 6-methoxygossypol and 6,6′-dimethoxygossypol

The triterpenoid aldehydes, gossypol (1), 6-methoxygossypol (2) and 6,6′-dimethoxygossypol (3); and the sesquiterpenoid aldehydes, hemigossypol (4) and methoxyhemigossypol (5), were isolated from 1-week-old roots of Gossypium hirsutum and G. barbadense and identified. This is the first report of 2 and 3 in nature and of 4 and 5 from healthy roots. Compounds 2 and 3 also constituted 30% of the total terpenoid aldehydes in the seeds of 1 cultivar of G. barbadense, but occurred only in trace quantities in those of G. hirsutum. Spectral data (UV, IR, NMR, MS) and proof of structure for 2 and 3 are presented.     

Development of a novel‐type transgenic cotton plant for control of cotton bollworm

The transgenic Bt cotton plant has been widely planted throughout the world for the control of cotton budworm Helicoverpa armigera (Hubner). However, a shift towards insect tolerance of Bt cotton is now apparent. In this study, the gene encoding neuropeptide F (NPF) was cloned from cotton budworm H. armigera, an important agricultural pest. The npf gene produces two splicing mRNA variants—npf1 and npf2 (with a 120‐bp segment inserted into the npf1 sequence). These are predicted to form the mature NPF1 and NPF2 peptides, and they were found to regulate feeding behaviour. Knock down of larval npf with dsNPF in vitro resulted in decreases of food consumption and body weight, and dsNPF also caused a decrease of glycogen and an increase of trehalose. Moreover, we produced transgenic tobacco plants transiently expressing dsNPF and transgenic cotton plants with stably expressed dsNPF. Results showed that H. armigera larvae fed on these transgenic plants or leaves had lower food consumption, body size and body weight compared to controls. These results indicate that NPF is important in the control of feeding of H. armigera and valuable for production of potential transgenic cotton.     

Relative consumption of three aphid species by the lacewing, Chrysoperla rufilabris, and effects on its development and survival

Relative consumption of three aphid species, Aphis gossypii Glover, Myzus persicae (Sulzer) and Lipaphis erysimi (Kaltenbach) (Homoptera: Aphididae), by larvae of the lacewing, Chrysoperla rufilabris (Burmeister) (Neuroptera: Chrysopidae), was determined in the laboratory, together with effects on lacewing development and survival. Percentages of survival of C. rufilabris from first instar to adult eclosion were significantly different among lacewing larvae fed different aphid species. When larvae were fed A. gossypii and M. persicae, all larvae developed to adulthood. All larvae died prematurely when they were fed L. erysimi. Developmental duration of C. rufilabris larvae was significantly shorter when larvae were fed A. gossypii (18.0 d) than when larvae were fed M. persicae (19.2 d). The number of fourth instar aphids consumed during development by C. rufilabris larvae differed significantly among individuals fed different aphid species. Chrysoperla rufilabris consumed an average of 168 M. persicae, followed by 141.6 A. gossypii, and only 26.6 L. erysimi. The percentage of these total number of aphids consumed by each larval stadium of C. rufilabris varied significantly among aphid species. The percentage of A. gossypii consumed by each larval stadium was similar to that for M. persicae, 12.1 and 11.4% by the first instar, 15.7 and 13.1% by the second instar, and 72.2 and 75.5% by the third instar, respectively; whereas in the case of L. erysimi, 23.3% of the total number of aphids were consumed by the first instar, 30.1% by the second instar, and 46.6% by the third instar.     

GhL1L1 affects cell fate specification by regulating GhPIN1‐mediated auxin distribution

Auxin is as an efficient initiator and regulator of cell fate during somatic embryogenesis (SE), but the molecular mechanisms and regulating networks of this process are not well understood. In this report, we analysed SE process induced by Leafy cotyledon1‐like 1 (GhL1L1), a NF‐YB subfamily gene specifically expressed in embryonic tissues in cotton. We also identified the target gene of GhL1L1, and its role in auxin distribution and cell fate specification during embryonic development was analysed. Overexpression of GhL1L1 accelerated embryonic cell formation, associated with an increased concentration of IAA in embryogenic calluses (ECs) and in the shoot apical meristem, corresponding to altered expression of the auxin transport gene GhPIN1. By contrast, GhL1L1‐deficient explants showed retarded embryonic cell formation, and the concentration of IAA was decreased in GhL1L1‐deficient ECs. Disruption of auxin distribution accelerated the specification of embryonic cell fate together with regulation of GhPIN1. Furthermore, we showed that PHOSPHATASE 2AA2 (GhPP2AA2) was activated by GhL1L1 through targeting the G‐box of its promoter, hence regulating the activity of GhPIN1 protein. Our results indicate that GhL1L1 functions as a key regulator in auxin distribution to regulate cell fate specification in cotton and contribute to the understanding of the complex process of SE in plant species.     

Impact of differential DNA methylation on transgene expression in cotton (Gossypium hirsutum L.) events generated by targeted sequence insertion

Targeted Genome Optimization (TGO) using site‐specific nucleases to introduce a DNA double‐strand break (DSB) at a specific target locus has broadened the options available to breeders for generation and combination of multiple traits. The use of targeted DNA cleavage in combination with homologous recombination (HR)‐mediated repair, enabled the precise targeted insertion of additional trait genes (2mepsps, hppd, axmi115) at a pre‐existing transgenic locus in cotton. Here we describe the expression and epigenome analyses of cotton Targeted Sequence Insertion (TSI) events over generations. In a subset of events, we observed variability in the level of transgene (hppd, axmi115) expression between independent but genetically identical TSI events. Transgene expression could also be differential within single events and variable over generations. This expression variability and silencing occurred independently of the transgene sequence and could be attributed to DNA methylation that was further linked to different DNA methylation mechanisms. The trigger(s) of transgene DNA methylation remains elusive but we hypothesize that targeted DSB induction and repair could be a potential trigger for DNA methylation.