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Ying Gao, Hao Liu, and Dong Pei

Monoclonal anti-indole acetic acid antibodies were used to monitor the temporal and spatial pattern of auxin during staminate flower differentiation in walnut (Juglans regia) cultivars Liaoning 1 and Liaoning 3. The relationship between morphological characteristics and histological structure was established. Seven stages of differentiation were recognized based on the visibility and color of the squama, bract, perianth, and rachis as follows: Stage 1, several bract primordia were present in the squama with catkins protruding from the squama as the only externally visible portion of the floret (Stage 1); the bract became externally visible, and the floret, perianth, and stamen primordia formed basipetally (Stage 2); the length of catkins were elongated, only bracts visible and getting brown (Stage 3); the bracts were brown and wrapped tightly, cellular specialization occurred to form a central core containing reproductive cells and tapetal cells that differentiated (Stage 4); the perianth became visible externally, reproductive cells and tapetal cells separated from the exterior layers of the anther wall (Stage 5); the anther walls were reduced to two cell layers (epidermis and endothecium) as the anthers became visible and matured (Stage 6); and the anther turned black, dehisced, and released its pollen grains (Stage 7). The histological differentiation of the flowers was related to auxin. The auxin signal was strongest in the shoot apical meristem (SAM) during bract primordia differentiation; thus, the SAM may be a site of auxin production. When the floral organs began centralized differentiation, auxin was distributed mainly in the differentiating tissues. Our findings indicate that a high level of auxin may strongly affect morphogenesis. Additionally, the tapetal and reproductive cells that arise during cellular specialization may be important for auxin production. The distribution of auxin was centralized in germ pores at the pollen grain surface, indicating that a high level of auxin induces pollen germination.

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Xin Hao, Yu Fu, Wei Zhao, Lifei Liu, Rengui Bade, Agula Hasi, and Jinfeng Hao

The MADS-box gene family encodes a type of transcription factor, and plays a key role in the growth and development of plants. Here, we identified 62 MADS-box genes in the melon (Cucumis melo) genome using bioinformatics methods. These genes were divided into type I Mα, Mγ, and Mδ subfamilies (26 members) and type II MIKCC subfamilies (36 members) by phylogenetic analysis. There were no genes in type II AGL12, BS, TM8, and MIKC* subfamilies, and type I Mβ subfamilies. Conserved motif analysis showed that all motifs had a subfamily-specific distribution except the M domain. The expression analysis of the MADS-box genes showed different expression characteristics. In summary, this study is the first to identify melon MADS-box genes and analyze their gene structures, subfamily distribution, and expression characteristics. These results provide a foundation for investigating the functions of the melon MADS-box genes.

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Yali Liu, Fuling Hao, Rui Meng, and Weirong Xu

The enzyme ACC oxidase (ACO), encoded by a small multigene family in many plants, catalyzes the terminal step in the ethylene biosynthesis pathway. In this research, based on the total RNA isolated from the flowers of Asia hybrids `Pollyanna' and Oriental hybrids `Sorbonne', we obtained two cDNA fragments of ACO genes (Genbank accession DQ062133 and DQ062134) by RT-PCR technique. The two cDNA fragments were reversely inserted into plant expression vector pWR306 respectively, and constructed two antisense ACO gene expression binary vectors harboring hygromycin phosphotransferase (hptII), glucuronidase (uid A), and a green fluorescent protein (GFP) gene in the T-DNA region. We have developed a system to produce transgenic plants in LiLium via Agrobacterium tumefaciens-mediated transformation of calli. Transformants were subjected to GFP expression analysis, PCR assay, and Southern hybridization to confirm gene integration.

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Ying Gao, Hao Liu, Ningguang Dong, and Dong Pei

We used anti-indole-3-acetic acid (IAA) monoclonal antibodies to monitor the temporal and spatial pattern of IAA during pistillate flower bud differentiation in the walnut (Juglans regia) cultivar Liaoning 1. Based on morphological changes, the process of pistillate flower bud differentiation was divided into five stages. The flower induction stage, which includes the early phase, midphase, and late phase, persisted from 25 Apr. to the end of May. The pedicel differentiation stage began on 5 June. The bract primordium stage began on 25 June and persisted through mid-March of the next year. Both the perianth and pistil differentiation stages persisted for nearly 2 weeks. During the floral induction period, little IAA was present in the shoot apical meristem (SAM); hence, the SAM may not always be a site of IAA production. IAA was obviously concentrated in cells of the first several layers of the SAM during pedicel primordium formation. High levels of IAA were also noted in the phyllome, young leaf tips, and vascular bundle of leaves and gemmae. This direct evidence indicates that no close relationship exists between IAA and physiological differentiation; instead, IAA may strongly affect morphogenesis. These findings comprise a first step toward elucidating the walnut flowering mechanism.

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Na Liu, Baoli Zhou, Xin Zhao, Bo Lu, Yixiu Li, and Jing Hao

Verticillium wilt (caused by Verticillium dahliae), a soilborne disease, often causes significant reductions of yield in eggplant (Solanum melongena L.) production where crop rotation is limited. Rootstock replacement through grafting is considered an effective method to control this disease. This 2-year study investigated the eggplant yield, resistance to verticillium wilt, and allelochemicals in root exudates of eggplant grafted onto a tomato rootstock. Both disease incidence and disease severity on grafted eggplant were markedly lower than those of nongrafted eggplants. Fifteen days after V. dahliae inoculation, grafted eggplants did not exhibit any infection, whereas the disease incidence and disease severity index of the nongrafted eggplants were 68.3% and 37.8% in 2006 and 66.7% and 36.3% in 2007, respectively. Twenty-five days after inoculation, disease incidences on grafted eggplants were only 8.1% and 9.5% in 2006 and 2007, respectively, but those of the nongrafted eggplants increased to 100%. As a result, early yield, total yield, and average fruit weight were significantly increased by grafting when inoculated with V. dahliae in 2006 and 2007. Mycelium growth of V. dahliae was inhibited by the root exudates of grafted eggplants. In contrast, the root exudates of nongrafted eggplants enhanced the mycelium growth. The gas chromatography–mass spectrometry analysis revealed that the composition in the root exudates released by grafted eggplants differed not only from the nongrafted eggplants, but also from the tomato rootstock plants. Ten chemical classes were isolated and identified in root exudates of grafted eggplants. Carbazoles, amines, azulene, and fluorene were only detected in the grafted eggplants. The relative contents of ester compounds were the highest in the root exudates from the grafted eggplant followed by derivatives of benzene, whereas the relative contents of benzene derivatives were much higher than that of the ester compounds in the root exudates from the nongrafted eggplant and tomato rootstock.

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Suxiao Hao, Yanfen Lu, Jing Liu, Yufen Bu, Qi Chen, Nan Ma, Zhiqin Zhou, and Yuncong Yao

Dwarfing rootstocks can improve the plant architecture of apple trees and increase production. Gibberellins (GAs) are crucial for plant growth and dwarfing traits. The receptor, GIBBERELLIN INSENSITIVE DWARF1 (GID1), plays an important role in the regulation pathway. However, the growth regulatory mechanism of GID1 in dwarf apple rootstock seedlings is not clear. In this study, we selected dwarf apple rootstock ‘SH6’ and its cross parents as materials to clone the GA receptor gene GID1c. There were two different sites in the alpha/beta hydrolase domain. The expression of GID1c in ‘SH6’ was lower than that in Malus domestica cv. Ralls Janet, with the decrease of GA content. We further conducted GA3 treatment and overexpression of GID1c in tissue culture seedlings of ‘SH6’, and the results showed that the expression of GID1c and biosynthesis genes increased and promoted the accumulation of hormone contents, which ultimately regulates the growth of ‘SH6’ dwarf apple rootstock seedlings. Our results suggest that GID1c may affect the plant architecture and dwarf traits of dwarfing rootstock and accelerate its application in orchards.

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Rui Zhang, Fang-Ren Peng, Dong-Liang Le, Zhuang-Zhuang Liu, Hai-Yang He, You-Wang Liang, Peng-Peng Tan, Ming-Zhuo Hao, and Yong-Rong Li

Scion wood of ‘Caddo’ and ‘Desirable’ pecan (Carya illinoinensis) was grafted onto the epicotyl of 1-month-old, open-pollinated ‘Shaoxing’ pecan seedlings for evaluation as a grafting technique to reduce the time to produce grafted trees. The results showed that seedlings grafted with “base scions” had higher survival than those grafted with “terminal scions” for both ‘Caddo’ and ‘Desirable’. Also, grafting with paraffinic tape could achieve greater success rate than that with medical tape. The most ideal time to perform this grafting was late April in Nanjing, China, when pecan seedlings were about 35 days old. This study demonstrated that the technique yielded successful epicotyl grafting of >70%, and it could thus be applied in practice.