Tamarixia radiata Waterson (Hymenoptera: Eulophidae) is a specific ectoparasitoid of the Asian citrus psyllid (PAC) Diaphorina citri Kuwayama (Hemiptera: Psyllidae), pest of economic importance in many citrus growing regions (^{Halbert and Manjunath, 2004}) by transmitting the great-negative bacteria known as Candidatus Liberibacter spp, causing the disease "Huanglongbing" (HLB) or citrus greening. (^{Garnier et al., 2000}; ^{Halbert and Manjunath, 2004}; ^{Bové, 2006}). Once the plant is infected and, depending on the age and condition of the crop, the HLB inevitably cause tree death in 5 to 10 years (^{Halbert and Núñez, 2004}; ^{Bové, 2006}).

An alternative to control the PAC, in areas where there is no management of this pest is T. radiata. This specific for D. citri (^{Aubert and Quilici, 1984}; ^{Zuparko et al., 2011}), it has been used in different countries in classical biological control programs with good results; Search high capacity at low host population, high fertility influenced by the density of the host (215-434 eggs/female) and its adaptability and synchronization with the plague would make a good candidate as a biological control agent to reduce phytophagous populations and consequently, it transmits disease (^{Chien et al., 1989}; ^{Chu and Chien, 1991}; ^{Chien, 1995}; ^{E´tienne et al., 2001}; ^{Hoy and Nguyen, 2001}; ^{Miranda et al., 2011}). Currently, the wasp is in Brazil, China, Guadeloupe, Indonesia, Mexico, Mauritius, Pakistan, the Philippines, Nepal, Taiwan, Vietnam, Puerto Rico and the United States of America (^{Chien et al., 1991}; ^{Waterston, 1922}; ^{Hoy and Nguyen, 2001}; ^{E'tienne et al., 2001}; ^{Halbert and Manjunath, 2004}; ^{Pluke et al, 2008}; ^{León and Setamou, 2010})

In Mexico there is a program for the control of D. citri based on the use of T. radiata as biological control agent (^{CNRCB, 2013}) and including the mass production of the parasitoid develops. T. radiata remains on nymphs of the Asian citrus psyllid, which in turn are produced in tender shoots of Murraya paniculata (L.) Jack (Rutaceae) host of D. citri. These plants are easy to use in greenhouses compared to other citrus species (^{Tsai, 2006}).

The main part of the production process of T. radiata is to have appropriate reproduction of the pest to infect host plants. The plants used must comply with the following requirements: preference of the pest, lots of foliage, lots of sprouting, fast growth, resistance to handling, ease of handling and resistance to pests and diseases. These requirements can only be met with proper fertilization, therefore, the present study aims to determine the best fertilizer to use on M. paniculata plants in order to get as much of outbreaks infested with D. citri that turn will give us a greater number of parasitoids.

This study was conducted in greenhouses of the National Reference Center for Biological Control (CNRCB) located on Highway Tecoman-station FFCC, 1.5 km from the Colonia Tepeyac in the City of Tecoman, Colima, Mexico.

Insects (PAC and T. radiata) as well as plants which are characterized by being of the same age and no pruning were provided by entomophagous area of CNRCB.

To develop the work is first made an apical and lateral pruning M. paniculata to stimulate sprouting. Pruning quickly, the products were applied directly to the soil to evaluate liquid except that Verdeabon compost and were applied as a solid product. Doses and evaluated products are listed in Table 1.

Table 1 Formulation and dosage of products used to stimulate sprouting of Murraya paniculata (L.) Jack (Rutaceae) in the greenhouses of the National Reference Center for Biological Control. Tecoman, Colima. 

Observations were performed every 24 h until five or more per plant shoots reach a size of between 5 and 10 cm. Outbreaks once achieved the desired length, the plants were placed in production cubes 70 x 70 x 70 cm mesh covered with anti-aphids be infested with a total of 100 males and 100 females of D. citri. After 7 days, the insects were removed with an oral sucker (25 mL vial). Observations were performed every 24 hours with the aid of a 10 x magnifier for the presence of eggs and once hatched, the emerged nymphs were grown up to the 3rd, 4th and 5th instar used in the production process of T. radiata plants, the presence of the required size nymphs were placed in buckets with the above characteristics and exposed to 75 parasitoid of T. radiata with a ratio of one male per two females over a period of 9 days. The temperature and relative humidity within the tilt buckets from 16.17 through 36.06 and 29.95 to 98.14 °C % RH (data logger Hobo^® Prov2 Ext Tem/RH onset).

Once the presence of meconium was observed in the plant, indicating parasitism of nymphs, the buds were cut and taken to the entomophages laboratory of CNRCB at a constant temperature of 27 °C. Buds and parasitized and non parasitized nymphs were counted with the aid of a stereoscope and subsequently placed for treatment in black cubes 40 x 40 x 40 cm and the upper concave and transparent part that allows the entry of light which stimulates the attraction of the parasitoid to emerge. The emerged parasitoids were collected and counted by treatment with an oral sucker 25 mL.

The experimental design of this study corresponded to a completely random design, where the unit undergoes plants of M. paniculata and response variables evaluated were: number of shoots per plant due to stimulation of the applied products, number of outbreaks infested by D. citri, number of nymphs present in each outbreak, number of nymphs parasitized outbreak T. radiata and quantity of parasitoids obtained by outbreak. Similarly, the percentage of parasitism and parasitoid emergence percent was recorded. The results were subjected to ANOVA test and if necessary means were separated by Tukey test (α= 0.05) (SAS 9, 2002).

It is known that the young shoots are ideal for oviposition substrate and development of the first nymphal instars of D. citri (^{Baños et al,, 2009a}; ^{Baños et al., 2009b}); however, during the present investigation it was observed that females equally psyllid may lay in mature buds or the underside of the leaf as presented in Algaenzims treatments and the control (Table 2).

Table 2 Effect of different products on the sprouting of Murraya paniculata (L.) Jack (Rutaceae) Infestation of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) and parasitism of Tamarixia radiata Waterson (Hymenoptera: Eulophidae). 

*Medias con la misma letra, estadísticamente son iguales (Tukey p< 0.05), ϻ= media, E.E= error estándar.

On the results to stimulate sprouting of M. paniculata by applying bio-stimulators can be observed graphically that three of the seven treatments were evaluated behave similarly in terms of the amount of buds, with the Zeagrow, and compost which Verdeabon greater number of outbreaks occur (Figure 1).

Figure 1 Number of shoots per plant produced by the implementation of six different bio-stimulators on Murraya paniculata (L.) Jack (Rutaceae). 

In the statistical analysis, there is a significant evidence with α= 0.05, which at least one treatment causes a different effect on the number of outbreaks issued by M. paniculata (F= 10.85, Pr <0.0001). It was observed that three treatments induce a similar effect, while the remaining four different forms behave. This can be seen by comparing Tukey where Zeagrow, Verdeabon and compost are the best treatments (Table 2).

There is little information on the implementation of growth bio-regulators on M. paniculata plants, ^{Baños et al. (2009b)} stated that, the use of these products at recommended doses, and the establishment of a periodic pruning, induce the plant to increased sprouting. Similarly, ^{Coleto (1994)} indicated that some phytoregulators induce cell elongation in shoots of fruit trees.

As regards the number of infested buds can be seen graphically nymphs of D. citri as many infested buds and compost Algaenzims unlike the remaining five treatments (Figure 2).

Figure 2 Number of outbreaks of Murraya paniculata (L.) Jack (Rutaceae) infested by the Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae). 

Data from the experiment with α= 0.05, statistically significant evidence shows that at least one treatment has a different effect on the amount of shoots infested with D. citri (F= 3.19, Pr <0.0085). This can be seen by comparing Tukey where Algaenzims and compost are those with the highest number of infested shoots (Table 2).

For the assessments of the number of nymphs per shoot (Figure 3), parasitized nymphs per shoot (Figure 4) and parasitoids obtained by bud (Figure 5), graphically compost is the best performing features. The statistical analysis shows no significant difference between the treatments (F= 0.69, Pr> 0.6550; F = 1.11, Pr> 0.3679; F= 1.31, Pr> 0.2672) (Table 2); however, for higher production of parasitoids in outbreaks of 5-10 cm, should take into account the number of outbreaks present in M. paniculata.

Figure 3 Number of nymphs of the Asian citrus psyllids Diaphorina citri Kuwayama (Hemiptera: Psyllidae) present for outbreak of Murrya paniculata (L.) Jack (Rutaceae). 

Figure 4 Number of nymphs of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) present for outbreak of Murraya paniculata (L.) Jack (Rutaceae) parasitized by Tamarixia radiata Waterson (Hymenoptera: Eulophidae). 

Figure 5 Number of parasitoids of Tamarixia radiata Waterson (Hymenoptera: Eulophidae) obtained on Diaphorina citri Kuwayama nymphs (Hemiptera: Psyllidae) produced by outbreak of Murraya paniculata (L.) Jack (Rutaceae). 

In the present study, we obtained a percentage of parasitism ranging from 64.67 to 96.13 percent with an emergence of the parasitoid T. radiata 70.08 to 95.23 (Table 3).

Table 3 Percentage of parasitism and emergence percent of the parasitoid Tamarixia radiata Waterson (Hymenoptera: Eulophidae) produced on nymphs Diaphorina citri Kuwayama (Hemiptera: Psyllidae) plants grown on Murraya paniculata (L.) Jack (Rutaceae). 

^{Qureshi et al. (2009)} mentioned that, the rates of parasitism of T. radiata during the spring and summer range in an average <20%, 39% in September and 56% in November in different parts of Florida USA. On the other hand, ^{Michaud (2004)} and ^{Qureshi and Stansly (2010)} indicated that, the rates of parasitism in Puerto Rico between January and April are 79-88%.

Predation levels, the use of insecticides, poor hibernation or inherent biological aspects of parasitoid that occurs naturally may be responsible for the different rates of parasitism of Florida and Puerto Rico compared with those obtained in the present study in greenhouses the CNRCB.