[Breeding and extension of Pseudostellaria heterophylla new variety "Shitai No.1" in Guizhou province].

Based on collections and researches of Pesudostellaria heterophylla germplasm resources from different areas of China, by using Shibing SB-4 provenance as materials, the new variety "Shitai No.1" was bred by mass selection, small plot variety comparative test, regional variety comparative test and field trial planting. Compared with "Qian taizishen No.1" and P. heterophylla land races. The disease and lodging resistance, root yield, polysaccharide content and the first grade rate of "Shitai No.1" have obvious advantages. In addition, it is relatively stable of yield in "Shitai No.1" in different places. It is demonstrated that "Shitai No.1" is a fine variety that adapt to the producing areas of P. heterophylla in Guizhou province, it is worthy to be promoted.

[Research on quality regionalization of cultivated Pseudostellaria heterophylla based on climate factors].

Maxent model was applied in the study to filtering the climate factors layer by layer. Polysaccharides and pseudostellarin B the two internal quality evaluation index were combined to analyse the interlinkages between climate factors and chemical constituents in order to search for the critical climate factors of Pseudostellaria heterophylla. Then based on the key climate factors to explicit the quality spatial distribution of P. heterophylla. The results showed that polysaccharides and climatic factors had no significant correlation, suggesting that the indicator was not climate-driven metabolites. Pseudostellarin B could construct regression model with the precipitation. And quality regionalization results showed that pseudostellarin B content presented firstly increased and then decreased trend from southeast to northwest, which was the consistent change with precipitation. It clearly proposed that precipitation was the key climate factor, which affected the accumulation of cyclopeptide compound for Pseudostellariae Radix.

[Quality evaluation and growing regionalization of Pseudostellaria heterophylla in Guizhou].

This paper is aimed to study the potential ecological suitability regionalization of Pseudostellaria heterophylla for selecting GAP planting base location and designing rational production layout. The ecological factors and contribution rate were determined by using maximum entropy (Maxent) model. Then, the information entropy theory was used to determine the relative importance of each environmental factor, and thus to determine the most limiting habitat criteria. Finally, the probable spatial distribution of P. heterophylla was determined based on GIS spatial analysis of habitat conditions. Meanwhile, the optimal index range of ecological factors was quantified. The moderately and highly suitable habitats were mainly located in Shibing, Huangping, Cengong, the middle and east of Kaili, the south of Yuqing, the west of Tongren. The percentage of moderately and highly suitable habitats for P. heterophylla in the study area was 3.64%, and its area was 6 405.39 km². The results also showed that seven dominant ecological factors controlled the distribution of P. heterophylla. These factors included agrotype, the warmest rain, aspect, slope, the warmest and highest temperature, contents of soil organic carbon, and the driest month precipitation. The habitat suitability assessment model based on GIS and Maxent model theory could accurately evaluate the habitat suitability distribution of P. heterophylla in Guizhou. In addition, we recommended Cengong and Zhenyuan county in Guizhou province as the worthy developing potential planting areas.

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Pseudostellaria heterophylla is a perennial herbaceous plant in the family Caryophyllaceae. The tuberous roots of P. heterophylla are highly valued in traditional Chinese medicine and have a high market demand. However, extended monoculture of P. heterophylla results in a significant decline in the biomass and quality, and escalates disease and pest problems. Therefore, it is important to understand the underlying mechanism and biocontrol methods for consecutive monoculture problems. With "Zheshen 2" as an experimental material, the changes in the contents of main nutrients in soil, phenolic acids and specific microbes under monoculture and different amendments were analyzed by using high performance liquid chromatography (HPLC) and qPCR. The results showed that consecutive monoculture of P. heterophylla led to a decrease in yield by 43.5% while the microbial fertilizer treatment and the paddy-upland rotation could relieve the consecutive monoculture problems. Available nitrogen, available phosphorus, available potassium and total potassium were significantly higher in the consecutively monocultured soils than in the newly planted soils. But consecutive monoculture resulted in soil acidification. HPLC analysis showed that conse-cutive monoculture of this plant did not lead to a consistent accumulation of soil phenolic acids. At middle stage of root expansion and at harvest stage, most of phenolic acids were even higher in the newly planted soils than in the consecutively monocultured soils. Furthermore, qPCR analysis showed that the amounts of three specific pathogens identified previously (i.e. Fusarium oxysporum, Talaromyces helicus, Kosakonia sacchari) were significantly higher in the consecutively monocultured soils than in the newly planted soils. However, the microbial fertilizer treatment and the paddy-upland rotation resulted in a significant decline in the population of these specific pathogens and improved the soil environment. In conclusion, the consecutive monoculture problems of P. heterophylla may be due to the rapid proliferation of host-specific pathogens, rather than the deficiency of soil nutrients and the autotoxicity of allelochemicals in root exudates. The results in this study could provide the theoretical basis to explore the underlying mechanism of replanting disease of P. heterophylla and its biocontrol strategies.

[Study on seed quality test and quality standard of Pesudostellaria heterophylla].

Referring to the rules for agricultural seed testing (GB /T 3543-1995) issued by China, the test of sampling, seed purity, weight per 1 000 seeds, seed moisture, seed viability and germination rate had been studied for screening seed quality test methods of Pesudostellaria heterophylla. The seed quality from different collection areas was measured. The results showed that at least 6.5 g seeds should be sampled and passed through 10-mesh sieve for purity analysis. The weight of 1 000 seeds was determined by using the 500-seed method. The phenotypic observation and size measurement were used for authenticity testing. The seed moisture was determined under the higher temperature (130 ± 2) degrees C for 5 hours. The seeds were dipped into 0.2% TTC sustaining 1 hour at 40 degrees C, then the viability could be determined. The break dormancy seeds were cultured on sand at 10 degrees C. K cluster analysis was applied for the data analysis, the seed quality from different collection areas grading of P. Heterophylla was described as three grades. The seed quality of each grade should reach following requirements: for first grade seeds, germination rate ≥ 86%, 1 000-grain weight ≥ 2.59 g, purity ≥ 87%, moisture ≤ 13.1%; for second grade seeds, germination rate ≥ 70%, 1 000-grain weight ≥ 2.40 g, purity ≥ 77%, moisture ≤ 14.3%; for third grade seeds, germination rate ≥ 41%, 1 000-grain weight ≥ 2.29 g, purity ≥ 76%, moisture ≤ 15.8%. The seed testing methods for quality items of P. heterophylla had been initially established, as well as the primary P. heterophylla seed quality classification standard.

Remnants of populations provide effective source material for reintroduction of an endangered Hawaiian plant, Schiedea kaalae (Caryophyllaceae).

PREMISE OF THE STUDY: Reintroductions may be essential to prevent extinction of many critically endangered species. Ideally, reintroduction efforts rely on adjacent source populations, but limited source material may necessitate crossing individuals from different and possibly distant populations. To determine the consequences of integrating multiple populations in reintroductions, we investigated levels of inbreeding depression, outbreeding depression, and heterosis for populations of Schiedea kaalae (Caryophyllaceae), an endangered species endemic to the Wai'anae and Ko'olau Mountains of O'ahu, Hawai'i. The possibility of gene flow among plants was explored through pollinator observations. METHODS: Individuals from ex situ living collections of nine populations were hand-pollinated with pollen from the same plant, plants from the same population (for three populations only), or plants from different populations. Progeny were outplanted into two common gardens, one in each mountain range on O'ahu. Cumulative fitness was estimated using several independent life history stages. KEY RESULTS: Inbreeding depression was minimal, and no outbreeding depression was detected. In contrast, strong heterosis was evident in progeny from between-population crosses, which had higher relative fitness than progeny from self-pollinations or within-population crosses. Observations of floral visitors provided the first evidence that biotic pollination may be important for this species. CONCLUSIONS: Results demonstrate the ability to conduct genetic rescue of rare species and suggest that reintroductions may be most successful using heterotic individuals from crosses between populations and at sites where pollinators are present and promote outcrossing.

[Determination and analyse of soil fertility of Pseudostellariae Radix planting base of Shibing County and Huangping County in Guizhou Province].

OBJECTIVE: To determine soil fertility of the Pseudostellariac Radix planting base of Shibing County and Huangping County in Guizhou Province, and to provide experimental basis for soil improvement and balanced fertilization. METHODS: 12 soil samples from Shibing County and 10 soil samples from Huangping County were involved in the detection. RESULTS: In the soil samples from Shibing County, the contents of organic matter, total N, available N, total P, available P and available K were higher, while the content of total P was at middle level. In the soil samples from Huangping County, the contents of total N, available P, total K and available K were higher,the contents of organic matter, total K and available N were at middle level. CONCLUSION: The level of soil fertility of the Pseudostellariae Radix planting base in Guizhou Province is common. The soil fertility of Shibing County is superior to that of Huangping County. The soil pH value of Shibing and Huangping County is low. In Shibing County, lime, farmyard manure and neutral fertilizer can be used to increase the pH of soil. Potash fertilizer can be increased, while nitrogen fertilizer and phosphate fertilizer can be reduced in the soil of Shibing County. Potash fertilizer and phosphate fertilizer will be increased, while nitrogen fertilizer can be reduced in the soil of Huangping County. The soil fertility can be improved by these measures.

[Breeding on eight strains of Pseudostellaria heterophylla based on phenotypic traits and quality in Guizhou province].

OBJECTIVE: To provide new germplasm materials for breeding new varieties of Pseudostellaria heterophylla. METHOD: The method of single plant selection was adopted, with the comparative experiments being carried out under the same conditions in Shibing county. The 8 plants of Shibing SB-4 were compared respectively with factor analysis for 27 phenotypic traits and 8 yield traits, and single factor variance analysis for the contents of polysaccharides. RESULT: Using factor analysis, 27 phenotypic traits were classified into 7 principal divisors and 8 yield traits were simplified into 3 principal divisors. The 4 strains of P. heterophylla, ZT-01, ZT-02, ZT-06 and ZT-07, performed better than others in the phenotypic traits, and ZT-01, ZT-02, ZT-03 and ZT-07 in the yield traits. The contents of polysaccharides of ZT-01, ZT-02, ZT-05 and ZT-08 showed significantly higher value. CONCLUSION: There is significant difference among the 8 strains of P. heterophylla in phenotypic traits, yield traits and quality traits, making it possible to select certain strains for different purposes. ZT-01 and ZT-02 can be breaded further. ZT-06 and ZT-07 were used as ornamental cultivars for its great phenotypic traits. ZT-03 with good resistance and high yield was taken as resistant variety, and ZT-05 would face next selection on the basis of its high content of polysaccharide.

[Characteristic research on soil nutrient of root of Pseudostellaria heterophylla root zone and non-root zone in different planting patterns].

OBJECTIVE: To research the influence of Pseudostellaria heterophylla planting patterns on soil nutrients. METHODS: The root zone and non-root zone soil nutrient status were analyzed, which included the crop rotation, intercropping, relay cropping, 1-year planting, 3-year planting, 6-year planting and 10-year planting. RESULTS: The results showed that there were differences in the soil nutrient of root zone among different planting patterns. The soil nutrient of root zone in seven different planting patterns were all significantly higher than that of non-root zone soil. And the soil nutrients of root zone showed a significant enrichment. The enrichment ratio of total K and Olsen K in 6-year and 10-year planting were both negative value, which indicated that the soil potassium was deficiency. The average pH value of root zone was 0.32 lower than that of non-root zone. There was a significant positive correlation between the organic matter value and the total nitrogen content in root zone and non-root zone (P < 0.01). The soil nutrients were high in crop rotation and intercropping, it would he helpful to improve soil quality and absorption to nutrient. CONCLUSION: The reasonable application of nitrogen fertilizer ,organic fertilizer and potash fertilizer can significantly increase the organic matter content in soil ,and it will help to retain the fertilizer capacity of soil.

[Seed dormancy mechanisms of Pseudostellaria heterophylla and its germination characteristics].

OBJECTIVE: Seed dormancy mechanisms, breaking dormancy and the optimal germinating conditions were studied to improve reproductive efficiency and quality standard of Pseudostellaria heterophylla. METHOD: The P. heterophylla seeds were stratified into wettish sand for 65 days under the temperature of -2-3 degrees C, embryo length and germination dynamic change of seeds were measured in prophase every other 10 days and in middle every other 5 days. The influences on seed germination and seedling growth under different temperatures, germination beds were investigated. RESULT: The P. heterophylla seeds embryo morphology was unchanged in stratification. The seeds could not germinate at stratification within the first 35 days. The germination rate and germination energy of seeds increased with the stratification time after 35 days. Germination rate, germination energy, plant height and fresh weight under the 10 degrees C were significantly higher than those under other temperatures. Germination rate, germination energy, plant height, fresh weight and dry weight of plant in sand bed were higher than those of plant in other beds, and seed mildew rate and abnormal seedling proportion of plant in sand bed were less than those of plant in other beds. CONCLUSION: The dormancy of P. heterophylla seeds is mainly the physiological post-maturation. The optimal germination conditions of seeds is 10 degrees C the culture of sand. The dormancy was broke under -2-3 degrees C of wettish sand for 45-50 days in practice.

[Study on tissue cultivation of adventitious roots of Pseudostellaria heterophylla].

OBJECTIVE: To systematically optimize the cultivation conditions of adventitious roots of Pseudostellaria heterophylla. METHOD: Tissue cultivation technology and ultraviolet spectrophotometry were adopted to observe the effect of inoculum volume, sucrose concentration, inorganic salt concentration, number of cultivation days, gradual scale-up cultivation and bubble different angles of bioreactor on the growth of adventitious roots of P. heterophylla, and determine the content of constituents such as saponin, polysaccharide and amino acid. RESULT: The propagation multiple of adventitious roots reached the maximum when the inoculum was 6 g in a 1 L culture shake flask. With the increase in sucrose concentration, the dry weight propagation multiples of adventitious roots followed an up and down trend. The inorganic salt concentration in a cultivation dish had a greater effect on the growth of adventitious roots, particularly 3/4 MS was the most favorable for the growth of adventitious roots. The growth curve of P. heterophylla was "S", with the biomass reaching the maximum at the 28th day. CONCLUSION: The inoculum volume, sucrose concentration, inorganic salt concentration, gradual scale-up cultivation and angles of bubble bioreactor had a significant effect on the growth of adventitious roots of P. heterophylla. The contents of saponin and amino acid in adventitious roots were higher than that in cultivated P. heterophylla, whereas the polysaccharide content were lower than that in cultivated P. heterophylla.

[Establishment of induction and culture system for hairy roots of Psammosilene tunicoides].

OBJECTIVE: To establish a culture system for Psammosilene tunicoides hairy roots, and provide technological aid for the large-scale production of P. tunicoides material. METHOD: The young leaves and stem segments of sterile plantlets were infected with ACCC10060 strain, and subsequently a culture system suitable for hairy roots growth was further established. RESULT: When explants were co-cultured with ACCC10060 (A600 0.8) on B5 media containing 20 mg x L(-1) Acetosyringo (AS) for 48 h, the hairy roots could be successfully induced, and it could achieve a higher induction rate using young leaves as explants than that of stem segments. The transfected hairy roots possessed the ability of kanamycin resistance and growth on hormone-free media, and synthesis of opines. All above results demonstrated that the present hairy roots originated in the infection of P. tunicoides tissues by ACCC10060 strains. After 35 d culture in liquid hormone-free MS (1/2 strength), the biomass of hairy roots increased 14.11 times (fresh weight) and 8. 39 times (dry weight), respectively, and the content of total saponins in hairy roots reached to 0.857% (DW), by contrast, it's only 0.388% and 0.217% in callus and seedlings respectively. CONCLUSION: Establishment of hairy roots culture of P. tunicoides provided a foundation for industrial production of active components from P. tunicoides culture.

[Observation on biological characteristics of wild Pseudostellaria heterophylla].

OBJECTIVE: To study the status and regular pattern in growing development of wild Pseudostellaria heterophylla. METHODS: Observed and studied on wild Pseudostellaria heterophylla during growth periods continuously. RESULTS: The growing development of Pseudostellaria heterophylla could be divided into three stages: asexual reproduction stage, coexistence of asexual and sexual reproduction stage. In the first two years of seed germination, there was only asexual reproduction, from which substitute tuberous root and cauline tuberous root were formed; Afterward flowering and fructifying sexual reproduction occurred to the plant that had developed from tuberous root while it continued with asexual reproduction; Only sexual reproduction existed during the last year of the Pseudostellaria heterophylla plant before its death. The Pseudostellaria heterophylla plant above ground grew from early spring to late autumn and over-wintered with tuberous root. There were two patterns of asexual reproduction: substitution with tuberous root in spring and expanding of adventitious root on stolon into cauline tuberous root in autumn. All tuberous roots of Pseudostellaria heterophylla were originated from adventitious root; The existence of tuberous root was one year at most; The rapidly expanding period of tuberous root was during February and March. CONCLUSIONS: The results provide a reference for reformative cultivation of Pseudostellaria heterophylla.

[Ribosomal DNA ITS sequence of analysis of Psammosilene tunicoides from different populations].

OBJECTIVE: To analyze the ribosomal ITS sequence variation of Psammosilene tuncolides W. C. Wu et C. Y. Wu from different populations, for identifying different local populations. METHODS: A pair of primers of 18SP1 and 26SP2 with PCR technique had been applied to study the ITS sequences. RESULTS: The sequences of ITS1, ITS2 and 5.8S are 225-229 bp, 166-170 bp and 261-264 bp. Among 12 local populations, the sequence of Kunming, Lijiang, Gejiu, Heqing of Yunnan and Yanyuan of Sichuan showed no variation, there were 1-4 variable sites (including 5.8S coding region) in pairwise comparison of the other 7 local populations, including Xuanwei, Huize, Zhongdian, Baoshan of Yunnan, Muli of Sichuan and Linzhi of Tibet. CONCLUSION: Comparative analysis shows that the ITS sequences of different local populations in the middle of Yunnan, southwest of Sichuan and west, northwest of Yunnan, southeast of Tibet, southwest of Sichuan have different fingerprint character, so the ITS sequences can be used to identify different local populations. The variation of ITS sequence of Psammosilene tunicoides is related to its geographical distribution.

[Growth comparison among Psammosilene tunicoides populations in tissue culture].

The comparison between the growth of eight populations from Psammosilene tunicoides at Yunnan Province was made by the tissue culture. The initial results showed out two populations from Yunshanping (Lijiang) and Xiaomoyu (Kunming) was dominant than orthers. It would be regard as one of fine germplasm resources for the culture of Psammosilence tunicoides.

[A research on fast-propagation of Pseudostellaria heterophylla].

The result of fast-propagation of Pseudostellaria heterophylla by tissue culture showed that the best explant was stem tip. It also showed the suitable inducement mediums were MS + 6-BA 2.0 mg/L + NAA 0.2 mg/L or MS + 6-BA 4.0 mg/L + NAA 0.4 mg/L for the clustered bud, MS + 2,4-D 4.0 mg/L + Kt 0.5 mg/L or MS + 2,4-D 3.0 mg/L + Kt 0.5 mg/L for the callus growth and 1/2 MS + NAA 0.3 mg/L for the root growth.