Supported by the Fundamental Research Funds for Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences (1630042017029; 1630042017008).

1 Introduction

In order to make full use of the under-forest resources of rubber plantations and improve the comprehensive benefits, the intercropping in the rubber plantation has become an important measure to improve the production efficiency of rubber[1], and it is also an effective method for rubber growers to get out of poverty[2]. The main modes of artificial rubber forest intercropping system include rubber-tea, rubber-sugar cane, rubber-pineapple, rubber-pepper, rubber-coffee, rubber-cinnamon, rubber-Fructus Amomi, rubber-banana, rubber-Alpiniaoxyphylla Miq. etc.[3-5]. Among them, the rubber-A. oxyphylla Miq. intercropping model is most widely developed, and has good economic and ecological benefits[6]. It has been popularized and promoted widely in Hainan, China. A. oxyphylla Miq. is a plant in the Zingiberaceae family, and its dried ripe fruit can strengthen the spleen and stomach, tonify the kidney and regulate the flow of vital energy. A. oxyphylla Miq. is mainly produced in Guangdong and Hainan, China, and it is also cultivated Fujian, Guangxi, Yunnan and other places[7]. A. oxyphylla Miq. is one of the Chinese well-known four "South Medicines", and it is also a variety contained in PharmacopoeiaofPeople’sRepublicofChina (2010 edition), with an application history of 1 300 years. The development of under-forest intercropping of A. oxyphylla Miq. can not only make up for the lack of market supply of A. oxyphylla Miq., but also meet the industrial demand of Hainan Province for vigorously developing the under-forest economy[3]. In the recent three years, the under-forest planting area of A. oxyphylla Miq. has been growing rapidly in Hainan. The planting area in 2013 was 4 648 ha, and the planting area in 2015 was 6 139 ha (HainanStatisticalYearbook). In 2016, Hainan Province further proposed the intensive development of forestry-medicine, fungus, livestock and poultry and other under-forest planting and breeding industries, and developed the under-forest economic area of 66 700 ha (including reclamation area of 30 000 ha)[8]. In accordance with the industrial development speed of A. oxyphylla Miq. in under-forest intercropping and the provincial plan requirements of developing under-forest economy in recent years, it is estimated that the development of the A. oxyphylla Miq. industry in 2017 will require about 7.5 million seedlings (calculated based on 2 250 plants per hectare, 3 333 ha for development). Therefore, it is particularly necessary to carry out the large-scale breeding of A. oxyphylla Miq. seedlings.

两年来，李莉一直很忙，却没有多大成就，工作稍微好点儿了而已，生活档次就是吃烤串掏腰包利索了点儿。存款嘛，四位数而已。至于许峰，一直在她的心尖荡漾。

管理上的复杂程度远远大于技术，而资源整合的难点恰恰可以通过BIM信息化技术来解决。充分考虑生产过程的要求，对构件的运输方案进行规划，如道路限高、桥梁荷载、施工堆场和构建转运做出合理的规划，及时传递信息，达到合理安排生产和施工工期，环环相扣，提高效率，降低成本的效果。

At present, Hainan Province is extensively promoting the under-forest planting of A. oxyphylla Miq., and the seedlings mainly adopt the traditional division propagation pattern. However, the cost of traditional division propagation is high[9], and excavating the ramets will affect the yield of the next season. The prolonged asexual reproduction will easily lead to population degeneration, poor plant resistance, quality deterioration of medicinal materials and other problems. The seed quantity of A. oxyphylla Miq. fruit is large, and each fruit contains about 25 seeds, very suitable for breeding seedlings. According to the investigation of research team, it is found that the flowering period of A. oxyphylla Miq. is long, about two months, and the inflorescence type of A. oxyphylla Miq. is raceme. In the whole flower axis, we can see the flowers at different levels of development, that is, the development of the flowers born at the base of the flower axis is early, while the development of the flowers born near the top of the flower shaft is late. The fruit at the base is mature first, and the fruit at the top is later ripened. And currently there is no uniform standard reference for the harvesting of A. oxyphylla Miq. seeds for raising seedlings. These factors cause great differences in the seed yield, thousand kernel weight, empty seed rate and number of grains per spike in different harvesting periods. There are few studies on the seed characteristics of A. oxyphylla Miq. Wu Zuqiang etal.[10] introduce the A. oxyphylla Miq. seed breeding technology, but it lacks the physiological parameters to quantify the harvesting standard of A. oxyphylla Miq. fruit. Qiu Hailian[11] uses the ISSR molecular markers technology to analyze the genetic traits of 68 A. oxyphylla Miq. germplasm resources in Danzhou, Qiongzhong, Baisha and other regions, and the genetic diversity of the germplasm resources is small. At the same time, it is found that the A. oxyphylla Miq. seed has the biological characteristics of long germination time and scattered germination[12]. Zeng Lin etal.[13] study the cryogenic temperature preservation method for A. oxyphylla Miq. seeds. It needs a lot of A. oxyphylla Miq. seeds to carry out the large-scale breeding of A. oxyphylla Miq. seedlings. The proper harvesting period of seeds is crucial to the quality of the seeds. If the harvesting is too early, the seeds are immature and the nutrient accumulation is insufficient; if the harvesting is too late, it is easy to fall or mildew, reducing the seed quality[14-15].

What time is the best harvest period of A. oxyphylla Miq. seeds? What kind of physiological properties are expressed from the A. oxyphylla Miq. fruits and seeds harvested in the best harvest period? These questions need further study. In this experiment, we studied the A. oxyphylla Miq. seed quality parameters and the biological characteristics of germination in different harvest periods, in order to explore the optimum harvest period of A. oxyphylla Miq. seeds, improve the germination rate of A. oxyphylla Miq. seeds, and provide the theoretical basis for the large-scale breeding.

2 Materials and methods

Seed viability is the potential ability of seeds to germinate or the viability of embryos. The TTC method was used to determine the seed viability index of A. oxyphylla Miq. in different harvest periods (Table 2), and it indicated that on the 80th d after flow-ering, the normal dyeing rate of seeds was only 22 %, and on the 100th d and 110th d after flowering, the normal dyeing rate of the harvested seeds reached 85% and 87% respectively. The results showed that the seed viability of A. oxyphylla Miq. harvested 100 d after the flowering of A. oxyphylla Miq. reached the maximum.

The seed moisture content (%)=(Weight of sample before drying-Weight of sample after drying)/Weight of sample before drying.

对照组：行NC化疗，即：①在患者住院的第1天、第8天行标准为25 mg/m的长春瑞滨静脉注射治疗；②住院的第1～5天行标准为25 mg/m的顺铂静脉注射治疗，连续用药60 d。

2.2 Methods

2.2.1 Determination of basic parameters of seeds. Determination of fruit dry/fresh weight ratio: first, 100 A. oxyphylla Miq. fruits were randomly selected in three groups, spread evenly over the sample box, placed in an oven at (105±2)℃ for 18 h of drying, taken out and immediately placed in a desiccator, cooled to room temperature and weighed. The fruit dry/fresh weight ratio is calculated as follows:

The fruit dry/fresh weight ratio=Dried sample weight/Sample weight before the drying.

Determination of thousand kernel weight: 1 000 seeds were randomly taken, and weighed, repeated three times.

2.2.3 Determination of the seed water absorption rate. The water absorption rate of seeds was measured by the weighing method. 500 A. oxyphylla Miq. seeds in different harvest periods were randomly selected and weighed, respectively, and these seed were soaked in distilled water at room temperature and placed in a thermostatic incubator at 27℃, measured at 1, 2, 4, 8 and 12 h, respectively. And then it was measured once every 24 h. During the determination, the seed surface water was first absorbed with the filter paper, and one hundred thousandth electronic balance was used for weighing the seeds. It was weighed for three times continuously until the seed weight was the same, that is, the water absorption was stopped. The water absorption rate is calculated as follows:

2.2.5 Observation of the structure and dynamic germination of A. oxyphylla Miq. seeds. 50 A. oxyphylla Miq. seeds were taken and soaked in water for 48 h, then sterilized with 3% hydrogen peroxide for 25 min. The treated seeds were placed in the culture dish (10 cm in diameter) with two layers of filter paper, and the right amount of water was added to be moist. It was cultured in the constant temperature incubator, and the temperature was controlled at (27±2)℃. During the period, the German SZX7 stereo microscope (1∶200) was used to observe the seed germination process, and it was pictured and recorded.

Picking method: During the flowering period of A. oxyphylla Miq., the flowering time of each plant inflorescence was listed and recorded, and the A. oxyphylla Miq. fruits were collected 80, 90, 100, 110 d after flowering, respectively, four groups collected each time (each group included 2 000 fruits).

2.2.2 Determination of the seed viability Using the TTC dyeing method, according to the biochemical (tetrazolium) method (TTC method) stipulated in GB/T 3543.7-1995 CropSeedTestingStandards-TestingofOtherItems, the seed viability was determined, with 100 seeds as one replication, repeated 12 times, and averaged. 1 200 A. oxyphylla Miq. seeds were placed in a triangular flask, filled with the distilled water to make seeds immersed in the liquid, and soaked for 24 h at room temperature. With the phosphate buffer with pH of 7.8, TTC was prepared into 1.0% solution, stored in a dark brown bottle. During the test, the seeds were cut in half along the central axis with a razor blade, placed in a 25 mL beaker, and a certain amount of 1.0% TTC solution was added to thoroughly soak embryos. Water bath temperature was maintained at 30℃ as constant temperature, and it stained for 3 h to observe the dyeing. It was counted according to the normal dyeing, partial dyeing but more than 1/2 of non-dyeing on the top of the radicle or cotyledons, full non-dyeing, respectively.

Determination of moisture content: According to InternationalSeedTestingStandard (International Seed Testing Association, 1985), 100 A. oxyphylla Miq. seeds were randomly selected in three parts, spread evenly over the sample box, placed in an oven and dried at (105±2)℃ for 18 h, taken out, immediately put into the drying dish, cooled to room temperature and then weighed. The moisture content is calculated as follows:

Water absorption rate (%)= (Wet weight-Dry weight)/Dry weight.

The two treatments were repeated three times, respectively, and the mean value was used to draw the seed water absorption curve.

3.1 Comparison of basic parameters ofAlpiniaoxyphylla Miq. fruits and seeds and seed viability in different harvest periods By observing and measuring the A. oxyphylla Miq. fruit color and dry/fresh weight ratio in four harvest periods (Table 1), it was found that 80-110 d after flowering, the A. oxyphylla Miq. peel color gradually turned from dark green to yellow-green; the dry/fresh weight ratio of A. oxyphylla Miq. fruit increased gradually and became relatively stable, and for the A. oxyphylla Miq. fruit harvested on the 100th d after flowering, the dry/fresh weight ratio was maintained in 0.345-0.358. The thousand kernel weight is an important indicator to reflect the seed size and fullness. As shown in Table 1, for the A. oxyphylla Miq. seed harvested on the 100th d and 110th d after flowering of A. oxyphylla Miq., the thousand kernel weight and seed moisture content reached a maximum value, and tended to be stabilized, and the thousand kernel weight and moisture content of A. oxyphylla Miq. seed were 11.075 g and 13.01%, respectively. The results showed that the fullness of A. oxyphylla Miq. seeds harvested 100 d after the flowering of A. oxyphylla Miq. was the best.

Germination rate (%)= (Number of seeds germinated ÷ Total number of sample seeds).

通过FMA评分判断，治疗前两组患者无明显差异，治疗后两组患者相比，差异有统计学意义（P＜0.05）。见表1。

2.3 Data analysis Data were analyzed and plotted using Excel 2003 and Origin Pro 8.1 software. The significant difference analysis was conducted on the data using DPS7.55 software (LSD method, P<0.05).

3 Results and analysis

2.2.4 Determination of the seed germination rate. 1 000 seeds in the four harvest periods were taken, respectively, and the mixture of clean sand (sifted by 10-mesh sieve) and seeds was evenly spread on the plastic sand tray (sand bed) padded with sand (plastic tray perforated at the bottom to ensure the excess water exudation). The sand bed thickness was 8 cm, then a layer of about 0.5 cm thick matured coconut bran was spread on the surface, and it was watered until the water at the bottom seeped. The plastic tray was placed in the incubator, and the water was evenly sprinkled with a spray to make the sand bed moist. The temperature was set to be 32℃ in the daytime, 24℃ in the night, and humidity was 80%. Four replicates were set up for each treatment. The germination rate was calculated daily and calculated for a total of 50 d. The germination rate is calculated as follows:

Table 1 The basic parameters ofAlpiniaoxyphylla Miq. fruits and seeds in different harvest periods

Harvesttime∥dPeelcolorchangesFruitdry/freshweightratioThousandkernelweightofseed∥gSeedmoisturecontent∥%80Darkgreen(0．291±0．012)d(8．664±0．021)c(11．09±0．32)c90Lightgreen(0．314±0．009)c(9．973±0．016)b(12．56±0．18)b100Yellow⁃green，asmallnumberofbrownspotsonthepeel(0．345±0．010)b(11．075±0．009)a(13．01±0．24)a110Yellow⁃green，brownspotsonmostofthepeel(0．358±0．008)a(11．088±0．011)a(13．09±0．30)a

Note: Lowercase letters indicate significant differences (P<0.05).

2.1 Materials Through the previous observation, it was found that farmers generally began to harvest the A. oxyphylla Miq. fruit 80 to 110 d after the flowering of A. oxyphylla Miq. The experiment was carried out in the A. oxyphylla Miq. demonstration base in Hongmao Town, Qiongzhong County, Hainan Province.

某日，一位旅客向火车站的上级单位投诉，说车站的服务员服务质量不好。事情的经过是：这位旅客坐的是软卧，在娄底车站下车，忘记了将自己的包拎上，他要求车站服务员帮其将包拿下来，服务员当时正忙着接车，委婉地拒绝了。这个旅客大发脾气，说自己坐软卧，是精品旅客，本来就多支付了票价，车站服务员理应给他提供类似服务。要求没有得到满足，他拨打了投诉电话。

Table 2 The seed viability ofAlpiniaoxyphylla Miq. in different harvest periods

Harvesttime∥dNormaldyeingPartlydyedbutthetopoftheradicleorcotyledonsnotmorethan1/2Notdyedatall802238409043342310085961108776

Note: Normal dyeing (seeds can grow into normal seedlings after germination); Partly dyed but the top of the radicle or cotyledons not more than 1/2 (abnormal development of seedlings); Not dyed at all (dead seeds).

3.2 Comparison of water absorption rate ofAlpiniaoxyphylla Miq. seeds in different harvest periods As can be seen from Fig.1, the A. oxyphylla Miq. seeds in different harvest periods showed different water absorption characteristics. Within 4 h before seed water absorption, the water absorption rate of A. oxyphylla Miq. seeds increased in 4 different harvest periods 80-110 d after flowering of A. oxyphylla Miq., up to 26.69%, 29.31%, 35.74% and 36.82% of total seed water absorption under different treatments, respectively; within 4-24 h, the growth of water absorption rate under different treatments slowed down, but there was still a lot of water absorption; within 24-72 h, the growth of water absorption rate slowed down; after 72 h, the seed water absorption under different treatments tended to stagnate, and it was close to the highest water absorption rate. Finally, the water absorption rate under different treatments was 13.90%, 16.49%, 23.59% and 23.71%, respectively. It indicated that the imbibition and water absorption stage for the A. oxyphylla Miq. seed germination needed about 72 d, and the water absorption rate of the seeds with late harvesting period was high.

3、专家咨询费管理的标准。专家咨询费是指临时受聘的专家所取得的费用，其标准按国家有关规定执行。专家咨询费在实际中争议较少，主要是专家咨询费的标准普遍不明确，存在较大随意性。同时，存在以虚构人员名单等方式虚报冒领或者套取专家咨询费的情况。

Fig.1 The dynamic seed water absorption ofAlpiniaoxyphylla Miq. in different harvest periods

3.3 Study on the seed germination rate ofAlpiniaoxyphylla Miq. in different harvest periods and dynamic observation of seed germination As can be seen from Fig.2, all the A. oxyphylla Miq. seeds in different harvest periods began to germinate after about 12 days of germination treatment, and the germination time was up to 50 d; with the delay of harvesting period, the germination rate of A. oxyphylla Miq. seeds increased gradually and became steady. The seed germination rate of A. oxyphylla Miq. harvested 80 d and 90 d after flowering was relatively low (P<0.05), and the seed germination rate increased slowly, and on the 50th d, the germination rate of in the 2 harvest periods was only 9.21% and 28.13%. And 37 d before the germination treatment of A. oxyphylla Miq. seeds, the seed germination rate of A. oxyphylla Miq. harvested 110 d after flowering was significantly higher than the seed germination rate of A. oxyphylla Miq. har-vested 100 d after flowering (P<0.05); 37 d after the germination treatment, there was a significant difference in the germination rate between the two harvest periods (P>0.05). On the 50th d, the germination rate in the two harvest periods reached 75.56%. The results showed that the harvest period was closely related to the seed germination rate, and the A. oxyphylla Miq. fruit was completely mature 100 d after flowering.

综艺节目、金庸作品大全、靓绝五台山或是电子竞技，我都不甚了解，只能默默看着朋友圈一波接一波的缅怀和庆祝。人们也许没有一期不落地看完《非常6+1》，没有精读过金庸的那副“对联”，但旧人旧物确是自己青春里实实在在的标点，一提到他们，仿佛就闪回到阳光灿烂的日子。

（1）硫磺尾气吸收单元在改造后，双塔吸收工艺流程比改造前的单塔吸收工艺流程优势在于：正常生产过程中，更有利于有机硫的吸收；由于新增吸收塔II中溶剂GL-DS对有羰基硫（COS）具有更好选择吸收性能，COS硫脱除率在69.71%，这是降低尾气SO2的主要因素。

Fig.2 Dynamic seed germination ofAlpiniaoxyphylla Miq. in different harvest periods

The A. oxyphylla Miq. seeds consisted of seed coat, embryo and endosperm, and the seed coat was fibrous, with micropyle at the top (Fig.3a). The micropyle was opposite to embryo, and it pierced the opening during germination. On the 4th d of seed germination treatment, the A. oxyphylla Miq. seed embryo expanded, that is, it began to bud (Fig.3b). On the 12th d, the A. oxyphylla Miq. seed embryo pierced the micropyle at the top of seed coat to germinate (Fig.3c and Fig.3d). On the 15-17th d, the A. oxyphylla Miq. seed embryo gradually differentiated into the radicle and coleoptile (Fig.3e and Fig.3f). On the 19th d, the radicle of A. oxyphylla Miq. seeds differentiated into two seed roots to absorb water and nutrients (Fig.3g).

Note: a. form and structure of matured seed; b. vertical-section on the 4th d of seed germination; c. form and structure on the 12th d of seed germination; d. vertical-section on the 12th d of seed germination; e. form and structure on the 15th d of seed germination; f. form and structure on the 17th d of seed germination; g. form and structure on the 19th d of seed germination; h. embryo in differentiation on the 12th d of seed germination; i. embryo in differentiation on the 15th d of seed germination; scale of 500 μm.

Fig.3 Seed form and anatomical structure ofAlpiniaoxyphylla Miq.

4 Conclusions and discussions

In this experiment, the basic parameters and germination characteristics of A. oxyphylla Miq. seeds during different harvest periods were analyzed and compared. This study determined the optimum harvest period of A. oxyphylla Miq. seeds, clarified the biological characteristics of slow seed water absorption and long germination cycle, and concluded that the seed coat was the main factor leading to slow seed water absorption and long seed germination cycle. The quality of seeds directly affects the yield and quality of crops, and the high-vigor seeds have good growth advantage and production potential, which is of great significance to the development of agricultural production[16]. Wang Hualei etal.[17] studied the relationship between the harvest time and the color, size, weight or germination rate of the seeds, to determine the suitable harvest period of Camptothecaacuminata seeds. Studies have shown that the seed vigor is closely related to the seed maturity[18-19]. With the increase of seed maturity, the seed weight increases, and the seed dry weight reached a peak in the physiological maturity period when the seeds show the highest germination rate and viability. In this study, by comparing the basic parameters of A. oxyphylla Miq. fruit and seed and seed viability in different harvest periods, it was found that the A. oxyphylla Miq. fruit reached its full maturity 100 d after flowering, and at this moment, the A. oxyphylla Miq. seed with high quality could be obtained in harvesting fruit. Therefore, the harvesting standards of the mature A. oxyphylla Miq. fruit were as follows: The A. oxyphylla Miq. fruit skin turned from green to yellow (yellow-green), there were a small number of brown spots on the peel, and it was spicy enough; the fruit dry/fresh weight ratio was greater than 0.34, and the thousand kernel weight of seed reached more than 11 g; the dried seed moisture content was about 13.01%-13.09%; using TTC method, the seed viability index was measured to be 85%.

In the process of seed germination, water is an indispensable condition. Water absorption is the primary part of seed germination. The seed germination always begins from the increase of moisture content, seed imbibition and water absorption[20]. The seed water absorption can be divided into three stages: imbibition and water absorption stage; stagnant period of water absorption; active water absorption stage. In general, as for the seeds of plants, the water absorption of seeds was fast in the first stage. This stage mainly relies on the internal seed colloid for imbibition and water absorption, nothing to do with the metabolic activity of seeds. However, the seeds, with hard seed coat (sometimes including peel), and difficulty in imbibed germination, are called hard seeds (or hard-coated seeds, hard-shelled seeds). The higher the degree of hardiness, the longer the soaking time required for the seeds to reach the imbibition state[21]. Through the study of the dynamic water absorption of A. oxyphylla Miq. seeds, it was found that the imbibition and water absorption stage for the A. oxyphylla Miq. seed germination took a long time, about 72 h; the maximum water absorption rate was about 23.71% at the imbibition and water absorption stage, which indicated that the water permeability of A. oxyphylla Miq. seeds was poor. It was inferred that the A. oxyphylla Miq. seeds were hard seeds. The fiber of A. oxyphylla Miq. seed coat is the main obstacle to the water absorption of A. oxyphylla Miq. seed, and in the production, the seedlings can use the "equal amount of sand " for seed rubbing treatment to improve the permeability of the seed coat, promote the seed imbibition and water absorption, and further shorten the seed germination time.

The seed germination rate determination method is a simple and practical method to evaluate crop seed viability[22]. By studying the dynamic A. oxyphylla Miq. seed germination in different harvest periods, it indicated that the seed germination rate of A. oxyphylla Miq. harvested 100 d after flowering reached the maximum. Further, the optimum harvesting period of the seeds was verified from the perspective of A. oxyphylla Miq. seed germination rate. By the study on the dynamic A. oxyphylla Miq. seed germination, it was found that the A. oxyphylla Miq. seeds germinated for a long time, the mature A. oxyphylla Miq. seeds began to germinate on the 12th d, and the germination rate on the 50th d reached 75.56%, namely the maximum value. According to the observation of A. oxyphylla Miq. seed structure, it was speculated that the seed coat was one of the main reasons for low seed germination rate and long germination cycle of A. oxyphylla Miq. The seed coat structure of A. oxyphylla Miq. hindered the exchange of water and gas, but it needs further study on the mechanism of how the seed coat impeded the entry of water and gas into the A. oxyphylla Miq. seeds.

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